LIBRARY 

OF    THE 

UNIVERSITY  OF  CALIFORNIA. 


BIOLOGY 
R 
G 


m 


m  1 


IMMUNE    SERA 

H^MOLYSINS,  CYTOTOXINS,  AND 
PRECIPITINS 


BY 


PROF.  A.  JVASSERMANN,  M.D. 

UNIVERSITY   OF    BEKL1N 


AUTHORIZED     TRANSLATION 

BY 

CHARLES  BOLDUAN,  M.D. 


FIRST  EDITION 


NEW  YORK 

JOHN   WILEY   &    SONS 

LONDON  :  CHAPMAN  &  HALL,  LIMITED 

1904 


-  c 


Copyright,  1904 

BY 
CHARLES  BOLDUAN. 


ROBERT   PRUMMONT),    PRINTER,  NEW    YORK. 


AUTHOR'S   PREFACE, 


During  recent  years  that  part  of  biology  which 
concerns  itself  with  the  reactions  resulting  from  the 
injection  of  organic  constituents  of  one  animal 
into  the  body  of  another  has  been  worked  up  ex- 
perimentally with  great  enthusiasm.  This  field, 
which  in  the  beginning  seemed  to  possess  only  a 
purely  scientific  interest,  has  now  yielded  numer- 
ous analogies  to  the  results  obtained  in  the  experi- 
mental study  of  natural  and  artificial  immunity 
against  infectious  diseases.  Furthermore,  the  re- 
sults of  these  investigations  have  been  found  ap- 
plicable to  many  clinical  questions,  as  well  as  to 
certain  other  problems  of  every-day  life.  It  is 
with  pleasure,  therefore,  that  I  heed  the  request  of 
one  of  the  editors  of  these  "Clinical  Lectures" 
[von  Bergman]  to  present  these  highly  interesting 
results  to  the  medical  profession  at  large.  It  is  not 
my  purpose  to  give  the  details  of  the  many,  oft- 
times  complicated  experiments  undertaken  by  vari- 
ous authors  to  support  or  refute  different  theories. 
The  following  sketch  is  intended  rather  to  intro- 
duce my  colleagues  to  the  essentials  of  the  subject. 

A.  W. 


TRANSLATOR'S  PREFACE. 


The  subject  of  serum  diagnosis  and  therapy, 
already  grown  to  considerable  proportions,  is  con- 
stantly increasing  in  importance.  The  lack  in  our 
language  of  any  simple  and  concise  exposition  of 
the  subject  has  led  the  translator  to  make  this 
excellent  treatise  of  Prof.  A.  Wassermann  more 
readily  accessible  to  the  English-reading  medical 
public.  The  presentation  of  the  subject  follows 
the  author's  course  of  lectures  given  at  the  Uni- 
versity of  Berlin  in  1903.  No  changes  whatever 
have  been  made  in  the  translation.  A  table  of 
contents  has,  however,  been  added. 

C.  B. 

BROOKLYN,  January,  1904. 


CONTENTS. 


ILEMOLYSINS. 

PAGE 

DEFINITION i 

NATURE  OF  THE  H^MOLYTIC  SERUM 3 

Active  and  inactive  sera 3 

Alexin  and  substance  sensibilatrice  of  Bordet 4 

Role  of  the  substance  sensibilatrice 4 

The  exciting  agent 6 

ARTIFICIAL  IMMUNITY  AGAINST  BACTERIA 7 

Bacteriolytic  power  of  serum 7 

Specific  bactericides 8 

Analogy    of    the    bacteriolytic     and    haemolytic 

processes 9 

Active  and  inactive  bacteriolytic  sera 9 

STUDIES    OF    EKRLICH    AND    MORGENROTH    ON    THE 

NATURE  OF  HAEMOLYSIS 10 

Complements  and  immune  body 15 

On  what  the  specificity  depends 17 

Difference  between  specific  and  normal  sera 18 

Diverging  views  of  Ehrlich  and  Bordet 18 

SIDE-CHAIN  THEORY  OF  EHRLICH 19 

Applied  to  the  production  of  antitoxins 20 

Toxins:    made  up  of  two  groups,  toxophore  and 

haptophore 20 

Toxoids 20 

vii 


vin  CONTENTS. 

PAGE 

Functions  of  the  side-chains 20 

Receptors 22 

Weigert's  overproduction  theory 22 

Extension  of  the  theory  to  the  production  of  other 

anti-bodies 26 

NORMAL  SERUM 29 

Its  haemolytic  and  bacteriolytic  action 29 

Active  and  inactive  normal  serum 30 

Inter-bodies  and  complements 31 

Action  not  entirely  specific 32 

Multiplicity  of  the  active  substances 33 

Hsemagglutinins  of  normal  serum 34 

Nature  of  the  agglutinins 36 

Agglutinoids 37 

Purpose  of  agglutination 37 

DIFFERENCE  BETWEEN  A  NORMAL  SERUM  AND  A  SPE- 
CIFIC IMMUNE  SERUM 38 

Practical  application 38 

IMMUNE  BODY:  ITS  NATURE 41 

Made  up  of  partial  immune  bodies,  according  to 

Ehrlich 41 

MetchnikofTs  views 43 

Where  immune  body  is  formed 45 

ANTI-HJEMOLYSINS  :  THEIR  NATURE 45 

Anti-immune  body 45 

Anti-complement 47 

Auto-anticomplernent 49 

FLUCTUATIONS  IN  THE  AMOUNT  OF  THE  ACTIVE  SUB- 
STANCES IN  SERUM 51 

COMPLEMENTS 53 

Their  origin 53 

Structure  of  the  complements 54 

Complementoids 54 

ISOLYSINS,  AUTOLYSINS,  AND  ANTILYSINS 55 


CONTENTS.  ix 


CYTOTOXINS. 

PAGE 

DEFINITION 59 

CYTOTOXIN  FOR  LEUCOCYTES,  LEUCOTOXIN 59 

NATURE  OF  THE  CYTOTOXINS:  THEIR  TWO  PARTS 59 

ANTI-CYTOTOXIN 59 

CYTOTOXIN  FOR  SPERMATOZOA,  SPERMOTOXIN.     ANTI- 

SPERMOTOXIN 60 

CYTOTOXIN  FOR  EPITHELIAL  CELLS 61 

CYTOTOXIN  FOR  NERVE-TISSUE,  NEUROTOXIN 62 

PRACTICAL  APPLICATIONS  IN  LEPROSY 63 

PRECIPITINS. 

DEFINITION 65 

PRECIPITIN  FOR  CASEIN  OF  MILK,  LACTOSERUM 66 

OTHER  PRECIPITINS 66 

NATURE  OF  THE  PRECIPITINS 67 

PRACTICAL  APPLICATION 68 

Wassermann's  method  of  differentiating  albumins.  69 

(a)  Used  to  test  old  blood  stains 69 

(6)   Used  to  separate  different  kinds  of  albumins.  70 

(c)  Used  to  distinguish  horse-flesh  from  beef.  71 

ANTI-PRECIPITINS 71 

ISO-PRECIPITINS 71 

CONCLUSION. 

CLINICAL  APPLICATIONS  OF  IMMUNE  SERA 73 

DIAGNOSTIC    VALUE    OF     ISO-AGGLUTININS    AND    OF 

ISOLYSINS 74 


tLEMOLYSINS,  CYTOTOXINS,  AND 
PRECIPITINS. 


I.  H^MOLYSINS. 

IN  1898  Belfanti  and  Carbone  showed  that  if 
horses  were  injected  with  red  blood-cells  of  rab- 
bits, the  serum  thereafter  obtained  from  the  horses 
would  have  acquired  an  appreciable  toxicity  for 
rabbits.  Shortly  after  this  Bordet  published  a 
very  interesting  series  of  experiments.  He  showed 
that  the  serum  of  guinea-pigs  after  these  had  been 
injected  several  times  with  3  to  5  c.c.  of  defibrin- 
ated  rabbits'  blood  acquires  the  property  to  dis- 
solve rapidly  and  intensely,  in  a  test-tube,  the  red 
blood-cells  of  a  rabbit;  whereas  the  serum  of  a 
normal  guinea-pig  is  incapable  of  doing  this,  or 
does  it  in  only  a  slight  degree.  Bordet  could 
further  show  that  this  action  is  a  specific  one,  i.e., 
the  serum  of  animals  treated  with  rabbit  blood 
acquires  this  dissolving  property  only  for  the  red 
cells  of  rabbits,  not  for  those  of  any  other  species 


2  IMMUNE  SERA. 

of  animal.  For  the  latter,  such  a  serum  is  no  more 
strongly  solvent  than  the  serum  of  a  normal  animal. 
The  same  property  that  Bordet  had  demonstrated 
in  the  serum  of  guinea-pigs  treated  with  rabbit 
blood  could  now  be  shown  for  the  sera  of  all  ani- 
mal species  treated  with  blood-cells  of  a  different 
species.  We  can  formulate  this  as  follows:  The 
serum  of  animals,  species  A,  after  these  have  been 
injected  either  subcutaneously,  intraperitoneally, 
or  intravenously  with  erythrocytes  of  species  B, 
acquires  an  increased  solvent  action  for  erythro- 
cytes of  species  B,  and  only  for  this  species.*  It 
is  therefore  a  specific  action.  We  call  'this  hce- 
molysis,  and  the  substances  which  effect  the  solution 
of  the  red  cells,  h&molysins  or  hcemotoxins. 

At  about  the  same  time,  and  independently  of 
Bordet,  similar  experiments  with  similar  results 
were  published  by  Landsteiner  and  v.  Dungern. 
As  a  result  of  this  work,  the  acquired  toxicity  of 
horse  serum,  found  by  Belfanti  and  Carbone  when 
they  treated  horses  with  red  cells  of  rabbits,  was 
explained.  The  serum  of  the  horses  so  treated  had 
become  h&molytic  for  rabbit  blood,  and  therefore 
caused  a  solution  or  destruction  of  the  red  cells 
in  the  living  body  just  as  it  did  in  a  test-tube. 

Agglutinating  Power  of  Haemolytic  Serum. — As  a 
further  result  of  his  experiments  Bordet  showed 
that  in  this  haemolytic  serum  still  another  property 

*  We  shall  point  out  a  few  exceptions  later  on. 


HAEMOLYSINS.  3 

had  been  increased,  namely,  the  power  to  clump 
the  rabbit  blood-corpuscles.  This  so-called  agglut- 
ination of  the  red  cells  occurs  previous  to  their  solu- 
tion. The  increase  in  the  agglutinating  power  of 
the  haemolytic  serum  is  a  specific  one.  For  if  an 
animal,  species  A,  be  treated  with  blood  of  species 
B,  the  serum  derived  from  A  will  have  acquired 
an  agglutinating  power  which  differs  from  that  of 
normal  serum  of  A  in  one  very  important  particular, 
namely,  in  that  it  is  specifically  increased  with 
respect  to  the  red  cells  of  species  B  or  its  nearest 
biological  relatives. 

That  normal  serum  of  an  unrelated  species  pos- 
sesses the  power  to  clump  the  red  cells  of  many 
other  species  had  already  been  shown  by  Creite 
and  Landois;  and  this  clumping  is  not  to  be  con- 
founded with  rouleaux  formation.  However,  the 
single,  specific  increase  of  the  agglutinating  power 
with  respect  to  a  distinct  and  definite  species  of 
red  cell  by  treatment  with  these  cells  was  first 
demonstrated  by  Bordet. 

Nature  of  Haemolytic  Sera — Active  and  Inactive  Sera 
— The  Two  Parts  of  Haemolysins. — This  author  now 
turned  to  a  further  study  of  the  action  of  the 
haemolysins,  and  was  able  to  show  that  the  sol- 
vent power  of  the  specific  haemolysins  depended 
on  the  combined  action  of  two  constituents  of  the 
specific  serum.  When  the  fresh  haemolytic  serum 
was  warmed  for  half  an  hour  to  55°  C.,  it  lost  its 
power.  If  to  this  inactive  serum  a  very  small 


4  IMMUNE  SERA. 

amount  of  the  serum  of  a  normal  guinea-pig  was 
added  (a  serum  which  of  course  was  not  haemolytic 
for  rabbit  red  cells),  the  full  haemolytic  power  was 
restored  to  this  inactive  serum.  In  other  words, 
it  had  been  reactivated  by  this  addition. 

This  experiment  permits  of  only  one  conclusion, 
namely,  that  the  hasmolytic  action  of  the  specific 
haemolytic  serum  depends  on  two  substances.  One 
of  these  is  able  to  withstand  heating  to  55°  C.,  and 
is  contained  only  in  the  specific  serum.  The  other 
is  destroyed  by  heating  to  55°  C.  and  is  contained 
not  only  in  the  specific  haemolytic  serum,  but  also 
in  the  serum  of  normal  untreated  animals. 

Alexin  and  Substance  Sensibilatrice  of  Bordet — Role  of 
the  Substance  Sensibilatrice. — Previous  to  this  work 
of  Bordet,  and  especially  as  a  result  of  the  researches 
of  Buchner,  it  had  been  known  that  there  were  con- 
stituents of  normal  blood-serum  which  were  ac- 
tively destructive  to  corpuscular  elements,  bac- 
teria, and  other  cells  with  which  they  came  in  con- 
tact. These  substances  had  been  termed  alexins 
by  Buchner.  This  term  was  retained  by  Bordet 
to  designate  that  constituent  of  normal  serum 
which  did  not  withstand  heating  to  55°  C.,  and 
which  was  one  of  the  factors  in  the  haemolytic 
process.  The  other  substance,  which  was  found 
only  in  the  specific  serum  and  which  withstood 
heating  to  55°  C.,  he  termed  substance  Sensibilatrice. 

According  to  Bordet,  therefore,  the  substances 
required  for  haemolysis  are  the  substance  sensibila- 


f   UNlV-.-.S 


trice  of  the  specific  haemolytic  serum  and  the 
alexin  which  exists  even  in  normal  serum.  The 
action  of  these  two  substances  Bordet  explains  by 
assuming  that  the  red  cell  is  not  vulnerable  to  the 
alexin  ;  just  as,  for  example,  there  are  certain  sub- 
stances that  will  not  take  a  dye  without  the  previous 
action  of  a  mordant.  The  substance  sensibilatrice 
plays  the  role  of  mordant.  It  makes  the  blood- 
cells  vulnerable  to  the  alexin,  so  that  the  latter  can 
attack  the  cells  and  dissolve  them.  The  alexin  he 
regards  as  a  sort  of  ferment  body  with  digestive 
powers. 

Bordet  says  further,  that  the  substance  sensi- 
bilatrice sensitizes  the  blood-cells  not  only  for  the 
alexin  derived  from  the  serum  of  the  same  species 
as  that  from  which  it  (the  substance  sensibilatrice) 
is  derived,  but  sensitizes  such  cells  also  for  the 
alexins  of  normal  sera  of  other  species.  For  ex- 
ample, in  the  foregoing  experiment  of  Bordet,  the 
substance  sensibilatrice  derived  from  the  guinea- 
pig  by  treatment  with  rabbit  blood  sensitizes  the 
red  blood-cells  of  rabbits  not  only  for  the  alexin 
of  normal  guinea-pig  blood,  but  also  for  the  alex- 
ins of  other  normal  sera.  In  another  experiment 
this  author  showed  that  rabbit  red  cells  sensitized 
with  an  inactive  specific  haemolytic  serum  derived 
from  a  guinea-pig  would  dissolve  rapidly  on  the 
addition  of  normal  rabbit  blood.  Here,  then,  the 
rabbit  red  cells,  sensitized  (according  to  Bordet) 
by  the  substance  sensibilatrice  of  the  guinea-pig, 


6  IMMUNE  SERA. 

dissolve  on  the  addition  of  the  alexin  of  their  own 
serum. 

The  Exciting  Agent.' — If  we  now  seek  to  discover 
the  constituent  part  of  the  red  cell  which  in  the 
treatment  excites  in  the  animal  body  the  production 
of  the  specific  haemolysin,  we  find  this  to  be,  accord- 
ing to  Bordet  and  v.  Dungern,  the  stroma  of  the  red 
cells.  This  separated  from  the  cell  contents  and  in- 
jected into  animals  will  likewise  excite  the  produc- 
tion of  specific  haemolytic  serum.  In  opposition  to 
this,  Nolf  assumes  that  the  stroma  excites  the 
production  of  the  above-mentioned  agglutinins, 
and  that  the  production  of  the  substance  sensi- 
bilatrice  is  called  forth  by  the  contents  of  the  red 
cells. 

Re*sume*. — Reviewing  the  important  facts  we  have 
learned,  we  find  them  to  be  as  follows:  By  means 
of  the  treatment  of  one  species  of  animal  with  the 
red  cells  of  a  different  one,  the  serum  of  the  first 
species  acquires  an  uncommonly  increased  power 
to  dissolve  and  to  agglutinate  the  red  cells  of  the 
second  species.  This  increased  haemolytic  power 
shows  itself  not  only  in  vivo,  so  that  an  animal 
so  treated  is  able  to  cause  red  cells  injected 
into  it  rapidly  to  dissolve  and  disappear,  but  it 
shows  itself  also  in  vitro  when  the  serum  of  this 
animal  is  used.  The  process  consists  in  the  com- 
bined action  of  two  substances,  that  which  is  ex- 
cited in  response  to  the  injection,  the  substance 
sensibila trice,  and  the  alexin  of  normal  serum. 


H&MOLYSINS.  7 

Artificial  Immunity  against  Bacteria — Bacteriolytic 
Power  of  Serum. — This  specifically  increased  solvent 
action  for  foreign  corpuscular  elements  on  the 
part  of  sera  of  animals  previously  treated  with 
the  same,  could  not  fail  to  be  of  the  great- 
est interest  to  bacteriologists;  for  a  most  surpris- 
ing similarity  showed  itself  between  this  and  the 
well-known  facts  of  artificial  immunity  against 
bacteria  as  they  had  been  developed  by  R.  Pfeiffer. 
In  order  to  make  this  clear  to  the  reader,  I  must 
dwell  for  a  moment  on  this  subject  of  artificial 
immunity  against  bacteria — for  example,  against 
living  cholera  bacteria.  A  normal  guinea-pig  is  able 
to  kill  and  dissolve  a  number  of  living  cholera 
bacilli  if  these  be  injected  intraperitoneally.  The 
freshly  drawn  serum  of  the  animal  possesses  the 
same  power.  If  this  serum  be  heated  to  55°  C., 
or  if  serurn  be  used  that  has  stood  for  some  time 
(eight  to  ten  days),  this  property  will  have  been 
lost.  This  power  of  normal  serum  and  other  body 
juices  of  the  living  animal  to  dissolve  appreciable 
quantities  of  many  bacteria,  Buchner,  as  already 
stated,  ascribed  to  certain  constituents  of  normal 
serum  which  he  called  alexins. 

These  alexins  are  of  very  delicate  constitution, 
decomposing  when  heated  to  55°  C.,  or  spontane- 
ously when  kept  outside  of  the  animal  body.  If 
we  inject  into  a  guinea-pig  a  very  minute  not  fatal 
dose  of  cholera  bacilli,  one  which  the  animal  is  able 
by  means  of  its  alexins  to  overcome,  and  if  we  then 


8  IMMUNE  SERA. 

gradually  increase  the  dose  injected,  it  will  be  pos- 
sible after  a  time  to  inject  at  one  dose  an  amount 
of  cholera  bacilli  that  represents  many  times  an 
ordinary  fatal  dose.  If  from  this  animal  we  now 
withdraw  serum  and  inject  it  into  another  animal, 
we  find  that  this  serum,  even  in  such  small  amounts 
as  the  fractional  part  of  a  centigram  or  even  of  a 
milligram,  is  able  to  protect  the  second  animal 
against  living  cholera  bacilli.  Under  the  influence 
of  these  small  amounts  of  serum  of  the  treated  ani- 
mal, the  organism  of  the  untreated  animal  is  able 
to  dissolve  large  amounts  of  cholera  bacilli,  amounts 
which  would  otherwise  be  invariably  fatal.  This 
process,  as  R.  Pfeiffer  showed,  is  a  specific  one,  i.e., 
the  serum  of  the  guinea-pig  treated  with  cholera 
bacilli  transmits  an  increased  solvent  power  only 
for  cholera  bacilli,  but  not  for  any  other  species  of 
bacteria.  The  active  substance  of  such  a  bacterio- 
lytic  immune  serum  Pfeiffer  called  a  specific  bac- 
tericide.  If  we  allow  some  of  this  specific  cholera 
immune  serum  to  remain  for  some  time  outside  of 
the  body,  e.g.  in  a  bottle,  and  then  test  it  for 
solvent  properties  against  cholera  bacilli,  not  in  a 
living  body  but  in  a  test-tube,  we  shall  find  that  its 
power  is  almost  nil.  If  we  add  to  this  serum  in 
the  test-tube  some  fresh  peritoneal  exudate  or 
some  other  body  fluid,  such  as  serum  of  a  normal, 
untreated  guinea-pig,  as  Metchnikoff  first  did,  we 
find  that  this  serum  has  now  acquired  the  power 
to  rapidly  dissolve  cholera  bacilli  even  in  a  test-tube. 


H&MOLYSINS  9 

Bordet,  in  1896,  showed  that  in  order  for  the  spe- 
cific immune  serum  to  dissolve  bacilli  in  a  test-tube, 
it  is  unnecessary  to  add  fresh  normal  serum  or 
peritoneal  fluid;  but  that  immune  serum  freshly 
drawn  from  the  vein  is  able  even  under  these  cir- 
cumstances to  dissolve  the  bacilli. 

Analogy  of  the  Bacteriolytic  and  Haemolytic  Proc- 
esses— Active  and  Inactive  Bacteriolytic  Sera. — Now 
that  the  main  points  in  cholera  immunity  are 
clear  to  us,  the  close  analogy  between  this  and  the 
subject  of  haemolysis  is  apparent.  Just  as,  when 
immunizing  an  organism  against  cholera  bacilli 
the  organism  responds  with  an  increased  solvent 
power  for  those  bacteria,  so  does  the  organism 
respond  when  it  is  treated,  i.e.  immunized,  with 
red  cells  of  another  species,  by  increasing  the  solv- 
ent power  of  its  serum  for  those  particular  cells. 
Furthermore,  just  as  the  haemolytic  process  was 
seen  to  depend  on  the  combined  action  of  two  sub- 
stances, one  developed  in  the  haemolytic  serum, 
the  other  already  present  in  normal  serum,  so  also 
in  the  bactericidal  process  just  studied  there  are 
two  factors.  It  is  easy  to  understand,  therefore, 
what  formerly  was  not  at  all  clear,  why  a  specific 
bactericidal  serum  against  cholera,  typhoid,  or 
other  infectious  disease  should  not  act  in  a  test- 
tube  unless  there  had  first  been  added  some  normal 
serum  (according  to  Metchnikoff),  or  there  had 
been  employed  a  perfectly  fresh  serum  (according 
to  Bordet) :  simply  because  in  either  of  these 


io  IMMUNE  SERA 

ways  the  alexin  necessary  to  co-operate  with  the 
substance  sensibilatrice  is  introduced.  This  alexin 
no  longer  exists  in  the  immune  serum,  if  this  be 
not  perfectly  fresh,  for  we  have  seen  that  it  decom- 
poses either  on  warming  or  spontaneously  on 
standing.  A  bactericidal  serum,  therefore,  that 
has  stood  for  some  time  is  incapable  of  dissolving 
bacteria.  It  is  possible,  however,  to  make  an  old 
inactive  serum  again  capable  of  dissolving  bacteria 
in  vitro  by  adding  ailittle  fresh  alexin,  according 
to  the  suggestion  of  Metchnikoff.  In  other  words, 
it  is  thus  reactivated.  Another  obscure  point  was 
cleared  up  by  these  studies:  why  a  specific  bac- 
tericidal serum  which  is  inactive  in  vitro  should 
be  intensely  active  in  the  living  body.  This  is 
because  in  the  living  body  the  serum  finds  the  alexin 
necessary  for  its  working,  which  is  not  the  case  in 
the  test-tube  unless  fresh  normal  serum  be  added. 
We  see  from  all  this  that  even  the  first  experiments 
in  haemolysis  have  served  to  clear  up  a  number  of 
practical  points  in  an  important  branch  of  bacteri- 
ology. 

Ehrlich  and  Morgenroth  on  the  Nature  of  Haemol- 
ysis.— In  continuing  the  study  of  hsemolysines  we 
must  note  particularly  the  researches  of  Ehrlich 
and  Morgenroth.  These  authors  asked  themselves 
the  following  questions:  (i)  What  relation  does 
the  haemolytic  serum  or  its  two  active  components 
bear  to  the  cell  to  be  dissolved?  (2)  On  what 
does  the  specificity  cf  this  haemolytic  process  de- 


HMMOLYSINS.  II 

pend  ?  Ehrlich  was  led  to  these  researches  partic- 
ularly by  his  so-called  Side-chain  Theory,  which 
we  shall  examine  in  a  moment. 

He  made  his  experiments  with  a  haemolytic 
serum  that  had  been  derived  from  a  goat  treated 
with  the  red  cells  of  a  sheep.  This  serum,  there- 
fore, was  haemolytic  specifically  for  sheep  blood- 
cells;  i.e.,  it  had  increased  solvent  properties  ex- 
clusively for  sheep  blood-cells. 

Basing  his  reasoning  on  his  side-chain  theory, 
Ehrlich  argued  as  follows:  "If  the  hsemolysin  is 
able  to  exert  a  specific  solvent  action  on  sheep 
blood-cells,  then  either  of  its  two  factors,  the  sub- 
stance sensibilatrice  of  Bordet  or  the  alexin  of  nor- 
mal serum,  must  possess  a  specific  affinity  for  these 
red  cells.  It  must  be  possible  to  show  this  experi- 
mentally." Such  in  fact  is  the  case,  and  the  ex- 
periments devised  by  him  are  as  follows: 

Experiment  i. — Ehrlich  and  Morgenroth,  as  al- 
ready said,  experimented  with  a  serum  that  was 
specifically  haemolytic  for  sheep  blood-cells.  They 
made  this  inactive  by  heating  to  55°  C.,  so  that 
then  it  contained  only  the  substance  sensibilatrice. 
Next  they  added  a  sufficient  quantity  of  sheep 
red  cells,  and  after  a  time  centrifuged  the  mixture. 
They  were  now  able  to  show  that  the  red  cells  had 
combined  with  all  the  substance  sensibilatrice,  and 
that  the  supernatant  clear  liquid  was  free  from  the 
same.  In  order  to  prove  that  such  was  the  case 
they  proceeded  thus:  To  some  of  the  clear  centri- 


12  IMMUNE  SERA. 

fuged  fluid  they  added  more  sheep  red  cells;  and, 
in  order  to  reactivate  the  serum,  a  sufficient  amount 
of  alexin  in  the  form  of  normal  serum  was  also 
added.  The  red  cells,  however,  did  not  dissolve  — 
there  was  no  substance  sensibilatrice.  The  next 
point  to  prove  was  that  this  substance  had  actually 
combined  with  the  red  cells.  The  red  cells  which 
had  been  separated  by  the  centrifuge  were  mixed 
with  a  little  normal  salt  solution  after  freeing  them 
as  much  as  possible  from  fluid.  Then  a  little  alexin 
in  the  form  of  normal  serum  was  added.  After 
remaining  thus  for  two  hours  at  37°  C.  these  cells 
had  all  dissolved. 

In  this  experiment,  therefore,  the  red  cells  had 
combined  with  all  the  substance  sensibilatrice, 
entirely  freeing  the  serum  of  the  same.  That  the 
action  was  a  chemical  one  and  not  a  mere  absorp- 
tion was  shown  by  the  fact  that  red  blood-cells  of 
other  animals,  rabbits  or  goats  for  example,  exerted 
no  combining  power  at  all  when  used  instead  of 
the  sheep  cells  in  the  above  experiment.  The 
union  of  these  cells,  moreover,  is  such  a  firm  one 
that  repeated  washing  of  the  cells  with  normal  salt 
solution  does  not  break  it  up. 

The  second  important  question  solved  by  these 
authors  was  this:  What  relation  does  the  alexin 
bear  to  the  red  cells  ?  They  studied  this  .by  means 
of  a  series  of  experiments  similar  to  the  preceding. 

Experiment  2.  —  Sheep  blood  was  mixed  with 
normal,  i.e.  not  haemolytic,  goat  serum.  After  a 


HMMOLYSINS.  13 

time  the  mixture  was  centrifuged  and  the  two  por- 
tions tested  with  substance  sensibilatrice  to  deter- 
mine the  presence  of  alexin.  It  was  found  that  in 
this  case  the  red  cells  acted  quite  differently.  In 
direct  contrast  to  their  behavior  toward  the  sub- 
stance sensibilatrice  in  the  first  experiment,  they 
now  did  not  combine  with  even  the  smallest  por- 
tion of  alexin,  and  remained  absolutely  unchanged. 
Experiment  3. — The  third  series  of  experiments 
was  undertaken  to  show  what  relations  existed 
between  the  blood-cells  on  the  one  hand  and  the 
substance  sensibilatrice  and  the  alexin  on  the 
other,  when  both  were  present  at  the  same  time, 
and  not,  as  in  the  other  experiments,  when  they 
were  present  separately.  This  investigation  was 
complicated  by  the  fact  that  the  specific  immune 
serum  very  rapidly  dissolves  the  red  cells  for  which 
it  is  specific,  and  that  any  prolonged  contact  be- 
tween the  cells  and  the  serum,  in  order  to  effect 
binding  of  the  substance  sensibilatrice,  is  out  of 
the  ~  question.  Ehrlich  and  Morgenroth  found  that 
at  o°  C.  no  solution  of  the  red  cells  by  the  haemo- 
lytic  serum  takes  place.  They  therefore  mixed  some 
of  their  specific  haemolytic  serum  with  sheep 
blood-cells,  and  kept  this  mixture  at  o°-3°  C.  for 
several  hours.  No  solution  took  place.  They  now 
centrifuged  and  tested  both  the  sedimented  red 
cells  and  the  clear  supernatant  serum.  It  was 
found  that  at  the  temperature  o°-3°  C.  the  red 
cells  had  combined  with  all  of  the  substance  sen- 


*4  IMMUNE  SERA. 

sibilatrice,  but  had  left  the  alexin  practically  un- 
touched. 

It  still  remained  to  show  the  relation  of  these 
two  substances  to  the  red  cells  at  higher  temper- 
atures. At  37°-4o°  C.,  as  already  mentioned, 
haemolysis  occurs  rapidly,  beginning  usually  within 
fifteen  minutes.  It  was  possible,  therefore,  to 
leave  the  cells  and  serum  in  contact  for  not  over 
ten  minutes.  Then  the  mixture  was  centrifuged 
as  before.  The  sedimented  blood-cells  mixed  with 
normal  salt  solution  showed  haemolysis  of  a  moder- 
ate degree.  The  solution  became  complete  when 
a  little  normal  serum  was  added.  The  supernatent 
clear  fluid  separated  by  the  centrifuge  did  not  dis- 
solve sheep  red  cells.  On  the  addition,  however, 
of  substance  sensibilatrice  it  dissolved  them  com- 
pletely. 

So  far  as  concerns  the  technique  of  the  experi- 
ments, I  should  like  to  observe  that  the  addition 
of  red  cells  in  this  as  well  as  in  all  the  following 
experiments  was  always  in  the  form  of  a  5%  mix- 
ture or  suspension  in  .85%,  i.e.  isotonic,  salt  solu- 
tion. 

The  significance  of  the  last  of  the  above-cited 
experiments  is  at  once  apparent.  It  is  that  the 
substance  sensibilatrice  possesses  one  combining 
group  with  an  intense  affinity  (active  even  at  o°  C.) 
for  the  red  cell,  and  a  second  group  possessing  a 
weaker  affinity  fone  requiring  a  higher  temperature) 
for  the  alexin. 


H&MOLYSINS.  15 

Nomenclature. — In  place  of  the  name  substance 
sensibilatrice  Ehrlich  first  introduced  the  term 
immune  body,  later  on  he  called  it  the  amboceptor. 
In  the  following  pages  we  shall  use  the  term  immune 
body,  as  this  had  already  been  used  by  R.  Pfeiffer 
to  designate  the  same  substance  in  bactericidal 
serum.  Other  names  proposed  for  this  substance 
have  been  substance  fixatrice  by  Metchnikoff,  copula, 
desmon,  preparator  by  Miiller.  Instead  of  the  name 
alexin,  Ehrlich  now  uses  the  term  complement  in  order 
to  express  the  idea  that  this  body  completes  the 
action  of  the  immune  body. 

In  contrast  to  the  specific  affinity  which  the  red 
cells  possess  for  the  immune  body,  these  cells  pos- 
sess no  affinity  whatever  for  the  alexin,  as  has  been 
shown  by  the  second  of  Ehrlich's  experiments. 
The  alexin,  therefore,  possesses  no  combining  group 
which  can  attach  itself  directly  to  the  red  blood- 
cell.  It  acts  on  these  cells  only  through  an  inter- 
mediary, the  immune  body,  which  therefore  must 
possess  two  binding  groups,  one  which  attaches  to 
the  red  blood-cell  and  the  other  to  the  alexin  of 
normal  serum.  As  already  stated,  the  group 
which  attaches  to  the  red  blood-cell  possesses  a 
much  stronger  affinity  than  that  which  combines 
with  the  alexin.  This  follows  from  the  last  two 
experiments  of  Ehrlich  before  cited,  in  which  he 
showed  that  at  the  lower  temperature  and  with 
both  substances  present  with  the  blood-cells  only 
the  immune  body  combined  with  the  cells,  while 


16  IMMUNE  SERA. 

the  alexin  remained  uncombined.  At  the  higher 
temperature  the  alexin  also  exerted  its  affinity,  for 
then  the  red  cells  combined  with  all  the  immune 
body  and  with  part  of  the  alexin.  We  saw  that 
after  a  time  the  red  cells  partially  dissolved,  but 
that  complete  solution  occurred  only  after  some 
fresh  alexin  had  been  added.  This  showed  that 
although  the  red  cells  had  combined  with  all  the 
immune  body  necessary  for  their  solution,  they  had 
been  unable  to  bind  all  the  alexin  necessary.  We 
may  say,  therefore,  that  that  group  of  the  immune 
body  which  combines  with  the  red  cell  has  a  stronger 
affinity  than  that  which  combines  with  the 
alexin. 

Role  of  the  Immune  Body. — According  to  Ehrlich, 
then,  the  role  of  the  immune  body  consists  in  this, 
that  it  attaches  itself  to  the  red  cell  on  the  one  hand 
and  to  the  complement  on  the  other,  and  in  this  way 
brings  the  digestive  powers  of  the  latter  to  bear 
upon  the  cell,  the  complement  possessing  no  affin- 
ity for  the  red  cell.  Immune  body  and  comple- 
ment have  no  very  great  affinity  for  each  other. 
At  o°  C.  they  may  exist  in  serum  side  by  side,  and 
they  combine  only  at  higher  temperatures. 

The  amount  of  immune  body  which  combines 
with  the  red  cells  may  vary  greatly,  as  the  experi- 
ments of  Bordet  and  of  Ehrlich  clearly  show.  Some 
red  cells  combine  with  only  just  enough  immune 
body  to  effect  their  solution.  Others  are  able  to  so 
saturate  themselves  with  immune  body  that  they 


H^EMOLYSINS.  17 

may  have  a  hundred  times  the  amount  necessary 
for  their  solution. 

On  What  the  Specificity  Depends. — From  the  pre- 
ceding it  follows  that  the  specific  action  of  the 
haemolytic  sera,  and,  I  may  at  once  add,  of  the  bac- 
tericidal sera  also,  is  due  exclusively  to  the  immune 
body.  This  possesses  a  combining  group  which  is 
specific  for  the  cells  with  which  the  animal  was 
treated;  e.g.,  the  combining  group  of  an  immune 
body  produced  by  treatment  with  rabbit  blood 
will  fit  only  to  a  certain  group  in  the  blood-cells  of 
rabbits;  an  immune  body  produced  by  treatment 
with  chicken  blood  will  fit  only  to  parts  of  the  red 
cells  of  chickens;  one  produced  by  treating  an  ani- 
mal with  cholera  bacilli  will  fit  only  to  this  species 
of  bacteria  and  combine  only  with  the  members  of 
it.  Keeping  to  the  well-known  simile  of  Emil 
Fischer,  the  relation  is  like  that  between  lock  and 
key,  each  lock  being  fitted  only  by  a  particular 
key. 

To  repeat — for  the  point  is  of  the  greatest  im- 
portance— the  role  of  the  immune  body  consists  in 
tying  the  complements  of  normal  serum,  which 
have  no  affinity  for  the  red  cells  or  for  the  bacteria, 
indirectly  to  these  cells  so  that  their  solution  and 
digestion  may  be  effected  by  the  complements. 
In  other  words,  the  immune  body  serves  to  con- 
centrate on  the  corpuscular  element  to  be  dis- 
solved all  the  widely  distributed  complement  found 
in  normal  serum. 


1 8  IMMUNE  SERA. 

The  relation  existing  between  complement,  im- 
mune body  (i.e.,  amboceptor)-  and  erythrocyte  is 
shown  in  the  accompanying  figure  reproduced 
after  Levaditi,  a  pupil  of  Ehrlich. 

i.  n. 

— zymotoxic  group 


f 

1 


COMPLEMENT— 

— hatophore  group 
— complimentophile  gr. 
IMMUNE  BODY— 

:ytophile  group 
— receptor 

CEL: 


FlG.    I. 

Difference  between  a  Specific  Serum  and  a  Normal 
One. — The  difference,  then,  between  a  specific  hcemo- 
lytic  or  a  specific  bactericidal  serum  and  a  normal 
one  consists  in  this — that  the  specific  serum  contains 
an  immune  body  which  is  specific  for  a  certain  cellu- 
lar element  and  by  means  of  which  the  complement 
present  in  all  normal  serum  can  be  concentrated  on 
this  cellular  element  to  cause  its  solution.  We  shall 
return  to  this  subject  later. 

Diverging  Views  of  Ehrlich  and  Bordet. — Now  if 
we  recall  the  first  experiments  of  Bordet  and  his 
conclusions  respecting  the  manner  in  which  the 
factors  concerned  acted,  we  shall  at  once  see  how 
Ehrlich  and  Bordet  differ.  Bordet  assumes  that 
the  substance  sen sibilat rice  (the  immune  body) 
acts  as  a  kind  of  mordant  on  the  red  cells  or  bac- 


HMMOLYSINS.  19 

teria,  sensitizing  these  to  the  action  of  the  alexin 
(complement).  According  to  Ehrlich,  however,  the 
process  is  not  analogous  to  a  staining  process, 
but  follows  definite  laws  of  chemical  combination, 
there  being,  in  fact,  no  affinity  whatever  between 
the  complement  and  the  blood-cells  or  bacteria. 
Furthermore,  according  to  this  authority,  the  com- 
plement always  acts  only  through  the  mediation 
of  the  immune  body,  which  possesses  two  combining 
groups;  one,  the  cytophile  group,  combining  with 
the  cell  and  another,  the  complementophile  group, 
combining  with  the  complement.  Both  observers 
have  devised  a  series  of  ingenious  experiments  to 
support  their  views.  But  as  these  can  interest 
only  the  specialist,  I  shall  omit  their  discussion 
here.  For  such  details  the  original  articles  may 
be  consulted. 

Ehrlich's  Side-chain  Theory.— The  results  of  the 
experiments  made  by  Ehrlich  to  determine  the  rela- 
tion of  immune  body,  complement,  and  cell  to  one 
another  served  as  a  further  support  for  his  so- 
called  Side-chain  Theory.  This  he  had  formulated 
several  years  before  in  order  to  explain  the  pro- 
duction of  antitoxin  and  other  specific  anti-bodies. 
Because  of  the  great  importance  of  this  theory, 
which  laid  the  foundation  for  much  of  this  work 
and  which  to-day  occupies  an  important  place  in 
our  literature,  and  also  because  it  will  serve  to 
make  the  following  more  readily  comprehensible, 
it  will  be  well  to  devote  a  little  time  to  its  study. 


20  IMMUNE  SERA. 

Originally  the  side-chain  theory  was  applied  by 
Ehrlich  only  to  the  production  of  the  specific  anti- 
toxins, i.e.,  substances  in  the  blood  which  act  not 
only  on  the  living  bacteria  but  also  and  especially 
on  their  dissolved  toxins.  Later  on  he  extended 
it  so  as  to  apply  also  to  the  formation  of  specific 
bactericidal  and  hsemolytic  substances  in  the  serum 
of  animals  treated  with  living  bacteria  or  with 
animal  cells. 

Toxins,  their  Toxophore  and  Haptophore  Groups — 
Toxoids — Special  Function  of  the  Side-chains. — The 
basis  of  the  theory  is  the  fact  that  poison  and 
counter-poison,  toxin  and  antitoxin,  combine 
directly  in  any  given  quantity.  This  combination 
always  occurs  in  definite  proportions  following  the 
laws  of  chemical  combination;  and,  still  following 
those  laws,  is  slower  at  lower  temperatures  than 
at  higher,  stronger  in  concentrated  than  in  di- 
lute form.  Ehrlich  could  further  show  that  each 
poison  for  which  by  the  process  of  immunizing 
one  can  develop  a  counter-poison  possesses  two 
groups  which  are  concerned  in  the  combination 
with  the  counter-poison  or  antitoxin.  One  of 
these,  the  so-called  haptophore  group,  is  the  combin- 
ing group  proper;  the  other,  the  toxophore  group,  is 
the  carrier  of  the  poison.  A  poison  molecule,  there- 
fore, might  lose  the  one,  the  toxophore,  and  still  be 
capable  by  means  of  its  haptophore  group  of  com- 
bining with  antitoxin.  Such  a  modified  poison, 
which  because  of  the  loss  of  the  toxophore  group 


H&MOLYSINS.  21 

can  hardly  be  called  a  poison,  but  which  still 
possesses  the  power  to  combine  with  antitoxin, 
Ehrlich  calls  a  toxoid.  Toxoids  may  be  produced 
spontaneously  in  old  poisons  through  decomposi- 
tion of  the  poison  molecule,  or  they  may  be  pro- 
duced artificially  by  causing  certain  destructive 
agents  such  as  heat  or  chemicals  to  act  on  bacterial 
poisons.  The  toxophore  group  is  a  very  delicate 
one  and  much  more  readily  decomposed  than  the 
combining  (haptophore)  group.  Ehrlich  reasoned 
that  in  order  for  a  poison  to  be  toxic  to  an  organ- 
ism, i.e.,  in  order  that  the  toxophore  group  be  able 
to  act  destructively  on  a  cell,  it  is  necessary  for  the 
haptophore  group  of  the  poison  to  combine  with 
the  cell.  "In  every  living  cell,"  Ehrlich  says, 
"  there  must  exist  an  active  central  body  [Leistungs 
Kern]  and  a  number  of  other  chemical  groups  or 
side-chains.  These  groups  have  the  greatest  vari- 
ety of  function,  but  especially  those  of  nutrition 
and  assimilation." 

The  side-chains,  then,  according  to  this  author, 
are  able  to  combine  with  the  greatest  variety  of 
foreign  substances  and  convert  these  into  nourish- 
ment suitable  to  the  requirements  of  the  active 
central  body.  They  are  comparable  to  the  pseudo- 
podia  of  the  lower  animals,  which  engulf  food  par- 
ticles and  assimilate  the  same  for  the  immediate 
use  of  the  organism.  In  order  that  any  substance 
may  combine  with  these  side-chains  it  is  necessary 
that  certain  very  definite  relations  exist  between 


22  IMMUNE  SERA. 

the  combining  group  of  the  substance  and  that 
of  the  side-chain.  To  repeat  the  simile  we  used 
above,  the  relation  must  be  like  that  of  lock  and 
key,  i.e.,  the  two  groups  must  fit  accurately.  Hence 
not  every  substance  will  fit  all  the  side-chains  of 
an  organism.  It  will  combine  only  with  those  for 
which  it  possesses  a  fitting  group. 

Receptors — Weigert's  Overproduction  Theory. — This 
doctrine  of  the  chemistry  of  the  organism's  me- 
tabolism Ehrlich  applied  to  the  action  of  toxins 
and  antitoxins.  "The  toxin,"  he  said,  "can  act 
only  when  its  haptophore  group  happens  to  fit  to 
one  of  the  side-chains,"  or  receptors,  as  he  now  pre- 
fers to  call  them.  As  a  result  of  this  combination, 
the  toxophore  group  is  able  to  act  on  the  cell  and 
injure  it.  If  we  take  as  an  example  tetanus,  in 
which  all  the  symptoms  are  due  to  the  central  ner- 
vous system,  the  side-chain  theory  assumes  that 
the  haptophore  group  of  the  tetanus  poison  fits 
exactly  and  is  combined  with  the  side-chains  or 
receptors  of  the  central  nervous  system.  Other 
experiments,  which  we  will  not  reproduce  here, 
have  shown  us  unquestionably  that  the  action  of 
the  antitoxins  depends  on  the  fact  that  this  com- 
bines with  the  haptophore  group  of  the  poison  and 
so  satisfies  the  latter 's  affinity.  Ehrlich,  therefore, 
concluded  that  the  antitoxin  is  nothing  else  than 
the  side-chains  or  receptors  which  are  given  off  by 
the  cells  and  thrust  into  the  circulation.  The  way 
in  which  these  side-chains  or  receptors  are  thrust 


H&MOLYSINS.  23 

off  as  a  result  of  the  immunizing  process,  Ehrlich 
explains  by  means  of  Weigert's  Overproduction 
Theory.  Weigert,  by  his  studies  in  new  tissue  for- 
mation, had  shown  that  whenever  there  is  a  defi- 
ciency in  the  organism,  the  latter  when  replacing 
this  is  incapable  of  remaining  within  bounds,  al- 
ways producing  more  than  is  required.  Ehrlich 
points  out  that  owing  to  the  combination  of  the 
toxin  with  the  side-chain  of  a  cell,  these  side-chains 
are  practically  lost  to  the  cell ;  that  the  latter  now 
produces  new  side-chains  to  replace  this  loss,  but 
that  this  production  always  goes  so  far  as  to  make 
a  surplus  of  side-chains;  that  these  side-chains  are 
thrown  off  by  the  cell  as  unnecessary  ballast  and 
then  circulate  in  the  blood  as  antitoxin.  The  same 
substances,  therefore,  which  when  part  of  the  cell 
combine  with  the  haptophore  group  of  the  toxin, 
enabling  that  to  act  on  the  cell,  when  circulating 
free  in  the  blood  combine  with  and  satisfy  this  hap- 
tophore group  of  the  toxin  and  prevent  the  poison 
from  combining  with  and  damaging  the  cells  of  the 
organism. 

Using  Ehrlich's  comparison,  this  may  be  likened 
to  an  iron  bar  within  a  building  which  owing  to  its 
attraction  causes  the  lightning  to  strike  the  build- 
ing. The  same  iron  bar,  however,  acts  as  a  protec- 
tion for  the  building  if  it  be  placed  outside  of  the 
same.  Let  us  bear  this  comparison  in  mind  as 
we  glance  at  the  facts  in  tetanus.  This  is  the  easi- 
est disease  in  which  to  study  the  relation  of  the 


24  IMMUNE  SERA. 

various  substances  and  processes  one  to  another, 
as  all  of  the  symptoms  are  referable  to  one  organ, 
the  central  nervous  system.  In  this  disease  the 
antitoxin  against  the  tetanus  poison  must  consist 
of  side-chains  or  receptors  thrust  into  the  circula- 
tion by  the  cells  of  the  central  nervous  system. 


toxophore  group 


POISON    MOLECULE 


haptophore  group 
receptor 


The  action  of  the  antitoxin  then  would  be  this,  that 
these  free  receptors  combine  with  the  haptophore 
group  of  the  tetanus  poison  as  soon  as  this  reaches 
the  circulation,  and  thus  prevent  this  poison /from 
combining  with  and  injuring  the  cells  of  the  central 
nervous  system.  This  I  was  able  to  prove  experi- 
mentally by  showing  (i)  that  the  central  nervous 
system  of  most  animals  susceptible  to  tetanus  is 
able  to  combine  with  the  tetanus  poison  in  vitro; 


H&MOLYSINS.  25 

and  (2)  that  such  a  mixture  of  tetanus  poison  and 
normal  central  nervous  system  is  innocuous  to  an- 
imals; because  certain  substances  present  in  the 
central  nervous  system  combine  with  and  thus  sat- 
isfy the  affinity  of  the  haptophore  group  of  the 
poison.  This  of  course  prevents  the  latter  from 
combining  with  any  of  the  cells  of  the  organism. 

Organs  other  than  the  central  nervous  system  do 
not  possess  this  property  of  combining  with  tetanus 
poison,  just  as  the  central  nervous  system  is,  on 
the  contrary,  incapable  of  combining  with  diphthe- 
ria poison  which  clinically  does  not  show  any  pro- 
nounced affinity  for  the  central  nervous  system. 
This  combination,  then,  of  central  nervous  system 
and  tetanus  poison  is  a  specific  one  in  conformity 
with  the  side-chain  theory.  Furthermore  it  has 
long  been  known  that  it  is  possible  to  immunize 
animals  (and  so  produce  antitoxins)  with  toxoids, 
i.e.,  with  poisons  that  possess  only  a  haptophore 
group.  This  supports  the  view  of  Ehrlich  that  the 
essential  feature  of  antitoxin  formation  is  the  com- 
bination of  the  haptophore  group  of  the  poison 
with  certain  definite  parts — the  receptors — of  the 
cell.  Conversely,  poisons  or  cells  the  affinity  of 
whose  haptophore  group  has  previously  been  satis- 
fied are  unable  to  excite  the  production  of  any 
antitoxin — excite  any  immunity.  That  is  because 
they  are  no  longer  able  to  combine  with  receptors 
of  the  cells,  v.  Dungern,  for  example,  showed  that 
blood-cells  which  had  previously  been  saturated 


26  IMMUNE  SERA. 

with  their  immune  body — in  other  words,  whose 
haptophore  group  had  been  satisfied — were  unable 
to  excite  the  production  of  any  haemolysin  when 
injected  into  animals. 

All  of  the  specific  relations  which  we  have  seen 
exist  between  toxin  and  antitoxin,  Ehrlich  and 
Morgenroth  in  their  experiments  above  noted 
found  to  exist  also  between  immune  body  and 
the  specific  blood-cell.  The  immune  body  must 
therefore  possess  a  haptophore  group  which  fits 
exactly  to  certain  receptors  or  side-chains  of  the 
red  cells,  just  as  the  anti-body  according  to  the 
side-chain  theory  possesses  a  group  that  fits  ex- 
actly into  the  specific  combining  group — i.e.,  hap- 
tophore group — of  the  toxin  or  toxoid  used  for  ex- 
citing the  immunity. 

If,  for  example,  we  produce  a  haemolytic  serum 
specific  for  red  cells  of  a  rabbit  by  injecting  an 
animal  with  these  cells,  the  haptophore  groups  of 
this  serum,  i.e.,  the  free  side-chains  thrust  off,  must 
possess  specific  combining  relations  with  the  red 
cells  of  rabbits.  That  such  is  the  case  in  the  hae- 
molytic immune  serum  we  saw  from  the  experi- 
ments of  Ehrlich  and  Morgenroth. 

The  Theory  Applied  to  the  Production  of  Other  Anti- 
bodies.— In  consequence  of  all  this,  Ehrlich  wid- 
ened the  application  of  his  side-chain  theory  so  as 
to  include  not  only  the  production  of  antitoxin 
but  also  the  production  of  bactericidal,  haemolytic, 
and  other  immune  bodies.  He  expressed  this 


HMMOLYSINS.  27 

somewhat  as  follows:  //  any  substance,  be  it  toxin, 
ferment,  constituent  of  a  bacterial  or  animal  cell,  or 
of  animal  fluid,  possess  the  power  by  means  of  a 
fitting  haptophore  group  to  combine  with 


(receptors)  of  the  living  organism,  the  possibility  for 
the  overproduction  and  throwing  off  of'  these  recep- 
tors is  given,  i.e.,  the  possibility  to  produce  a  cor- 
responding anti-body. 

Specific  anti-bodies  in  the  serum  as  a  result  of 
immunizing  processes  can  only  be  produced,  there- 
fore, by  .  substances  which  possess  a  haptophore 
group  and  which,  in  consequence,  are  able  to  form  a 
firm  union  with  a  definite  part  of  the  living  organ- 
ism, the  receptor.  This  is  not  the  case  with  alka- 
loids, e.g.,  morphin,  strychnin,  etc.,  which  accord- 
ing to  Ehrlich  enter  into  a  loose  union,  a  kind  of 
solid  solution  with  the  cells.  It  is  for  this  reason 
that  we  are  unable  to  produce  any  anti-bodies  in 
the  blood  serum  against  these  poisons.  Ehrlich 
says  further  that  all  of  the  substances  taking  part 
in  the  production  of  immunity,  including  of  course 
complement  and  immune  body,  have  certain  defi- 
nite affinities  for  each  other,  and  in  order  to  act 
they  must  fit  stereochemically  to  each  other. 

As  we  have  already  seen,  we  are  able  by  means 
of  the  injection  of  a  variety  of  substances  or  cells 
to  produce  a  similar  variety  of  immune  bodies  in 
the  serum.  Thus  we  can  immunize  a  rabbit  so 
that  its  serum  will  possess  specific  haemolytic 
bodies  against  the  red  cells  of  guinea-pigs,  goats, 


28  IMMUNE  SERA. 

chickens  and  oxen  and  specific  bactericidal  bodies 
against  cholera  and  typhoid  bacilli,  etc.,  and  as  we 
shall  see,  still  other  groups  of  anti-bodies. 

Multiplicity  of  Complements. — Under  these  circum- 
stances an  important  question  presents  itself: 
Is  there  in  normal  serum  one  single  complement 
which  completes  the  action  of  all  these  various 
immune  bodies,  one,  for  example,  which  in  the 
above  illustration  will  fit  all  the  haemolytic  im- 
mune bodies  as  well  as  all  the  bactericidal  ones,  or 
are  there  a  great  many  different  complements? 
Ehrlich,  as  a  result  of  his  experimental  work  with 
Morgenroth,  claims  that  the  latter  is  the  case; 
namely,  that  it  takes  a  different  complement  to  fit 
the  immune  body  specifically  haemolytic  for  guinea- 
pig  blood  than  it  does  to  fit  that  specific  for 
chicken  blood. 

Bordet,  on  the  other  hand,  assuming  that  the 
immune  body  plays  the  role  of  mordant,  believes, 
as  does  also  Buchner,  that  there  is  but  one  single 
complement  in  the  serum.  According  to  him, 
this  complement  is  able  to  dissolve  blood-cells  as 
well  as  bacteria  after  these  have  been  sensitized 
by  their  specific  immune  body.  Each  of  these 
authors  supports  his  claims  by  means  of  ingenious 
experiments,  for  the  details  of  which,  however, 
we  must  refer  to  the  original  articles,  as  they  re- 
quire the  knowledge  of  a  specialist  for  their  com- 
prehension. As  a  result  of  my  own  work  I  accept 
Ehrlich's  view,  that  of  the  multiplicity  of  the  com- 


H&MOLYSINS.  29 

plements.  One  thing  at  least  I  regard  as  proven, 
that  the  complement  which  fits  to  the  bactericidal 
immune  body  is  different  from  that  for  the  hsemo- 
lytic  immune  body.  According  to  his  most  recent 
work  this  view  is  also  shared  by  Metchnikoff. 
Later  on  we  shall  see  that  this  is  not  merely  an 
academic  question,  but  one  of  great  practical  im- 
portance. 

Normal  Serum,  its  Haemolytic  and  Bacteriolytic 
Action. — Inquiring  now  into  the  essential  differ- 
ence between  a  specific  haemolytic  or  bactericidal 
serum  and  a  normal  one,  we  must  first  of  all  study 
the  behavior  of  normal  serum  toward  foreign  red 
cells  and  bacteria.  It  has  long  been  known  to 
physiologists  that  fresh  normal  serum  of  many 
animals  has  the  power  to  dissolve  blood-cells  of 
another  species.  This  was  studied  especially  by 
Landois.  One-half  to  one  c.c.  of  normal  goat  serum, 
for  example,  is  able  to  dissolve  5  c.c.  of  a  5%  mix- 
ture (in  normal  salt  solution)  of  rabbit  or  guinea- 
pig  red  cells.  In  the  same  .way  these  red  cells 
are  dissolved  by  the  sera  of  oxen,  of  dogs,  etc. 
This  normal  globulicidal  property  of  the  serum  cor- 
responds to  another  which  fresh  normal  serum  was 
found  to  possess,  namely,  the  property  to  dissolve 
appreciable  quantities  of  many  species  of  bacteria. 
This  analogy  was  pointed  out  by  Fodor,  Nutall, 
Nissen,  and  especially  by  Buchner.  We  call  this 
the  bactericidal  property  of  fresh  normal  serum. 
Buchner,  as  we  have  already  seen,  had  studied 


30  IMMUNE  SERA. 

this  carefully  and  ascribed  the  action  to  a  substance 
found  in  all  normal  serum,  which  he  called  alexin. 
According  to  his  experiments,  this  is  a  very  unstable 
substance,  decomposing  spontaneously  on  standing 
or  on  heating  for  a  few  minutes  to  55°  C.,  or  readily 
on  the  action  of  chemicals.  According  to  this 
author  all  the  globulicidal  and  bactericidal  func- 
tions of  normal  serum  are  performed  by  this  one 
substance,  the  alexin. 

Active  and  Inactive  Normal  Serum. — Ehrlich  and 
Morgenroth  now  took  up  the  study  of  the  hae- 
molytic  action  of  normal  serum.  They  sought 
particularly  to  discover  whether  in  normal  serum 
the  haemolytic  property  depended  on  the  action  of 
a  single  substance,  the  complement  (Buchner's 
alexin),  or  whether  here  as  in  the  specific  hsemo- 
lytic  serum  it  depended  on  the  combined  action 
of  two  substances.  For  this  purpose  they  used 
guinea-pig  blood,  which  '  is  dissolved  by  normal 
dog  serum.  If  this  serum  was  heated  to  55°  C.,  it 
lost  its  haemolytic  power.  It  was  necessary  now  to 
show  that  in  this  inactive  dog  serum  there  re- 
mained a  second  substance  which  could  be  reacti- 
vated after  the  manner  of  reactivating  an  old  spe- 
cific haemolytic  serum.  This  had  its  difficulties, 
for  they  could  not  add  normal  dog  serum.  This, 
as  we  saw,  is  already  haemolytic  for  guinea-pig 
blood.  "  Possibly,"  said  they,  "  there  exists  a  com- 
plement of  another  animal  which  will  fit  the  hypo- 
thetical second  substance  of  this  dog  serum." 


H&MOLYSINS.  31 

This  proved  to  be  the  case,  the  complement  of 
guinea-pig  blood  fulfilling  the  requirements.  If 
they  added  to  the  inactive  normal  dog  serum  about 
2  c.c.  normal  guinea-pig  serum,  the  haemolytic  prop- 
erty was  restored  and  the  guinea-pig  red  cells 
dissolved  completely.  This  can  only  be  explained 
by  assuming  that  in  guinea-pig  blood  there  exists 
a  complement  which  happens  to  fit  the  hapto- 
phore  group  of  the  second  substance,  or  inter-body, 
of  the  normal  dog  serum.  This  combination  of 
guinea-pig  blood,  inactive  normal  dog  serum, 
and  a  reactivating  normal  guinea-pig  serum  is  the 
best  possible  one  to  demonstrate  the  existence  in 
normal  dog  serum  of  an  inter-body ;  for  the  guinea- 
pig  serum  should  be  the  best  possible  preservative 
for  the  guinea-pig  red  cells.  The  haemolysis  fol- 
lowing the  addition  of  this  serum  shows  positively 
the  existence  of  a  substance  in  the  dog  serum 
which  has  acted  with  something  in  the  guinea-pig 
serum.* 

Inter-body  and  Complement. — We  see,  then,  that 
the  haemolytic  action  of  normal  sera  depends,  just 

*  Of  such  combinations,  i.e.,  combinations  in  which  a 
complement  derived  from  the  same  animal  from  which  the 
red  cells  are  derived  fits  to  the  inter-body  of  other  species 
of  animals,  causing  the  solution  of  red  cells  of  the  latter, 
Ehrlich  and  Morgenroth  found  still  other  examples.  For 
instance,  guinea-pig  blood,  inactive  calf  serum,  guinea-pig 
serum;  goat  blood,  inactive  rabbit  blood,  goat  serum; 
sheep  blood,  inactive  rabbit  blood,  sheep  serum ;  guinea-pig 
blood,  inactive  sheep  serum,  guinea-pig  serum. 


32  IMMUNE  SERA. 

as  that  of  the  specific  haemolytic  sera,  on  the  com- 
bined action  of  two  bodies:  one,  the  inter-body, 
which  corresponds  to  the  immune  body  of  the  spe- 
cific sera,  and  a  second  or  complement.  In  speaking 
of  the  constituents  of  normal  serum,  Ehrlich  and 
Morgenroth  prefer  to  ,use  this  term  inter-body  to 
distinguish  it  from  the  immune  bodies  of  specific 
haemolytic  sera. 

Action  Not  Entirely  Specific. — It  has  also  been 
found  that  there  frequently  exist  normal  sera  which 
are  haemolytic  not  only  for  one  species  of  red  cell 
but  for  several.  We  saw,  for  instance,  that  normal 
goat  serum  dissolved  the  red  cells  of  guinea-pigs 
and  rabbits.  The  question  now  arises,  Is  this  prop- 
erty of  normal  goat  serum  due  to  two  inter- 
bodies  existing  in  the  serum  side  by  side,  one  fitting 
the  red  cells  of  the  guinea-pig,  the  other  those  of 
the  rabbit?  Ehrlich  and  Morgenroth  answered 
this  in  the  affirmative,  for  in  the  following  experi- 
ment they  succeeded  in  having  each  of  the  two 
inter-bodies  combine  with  its  respective  cell.  To 
some  inactive  normal  goat  serum  they  added  rab- 
bit blood  and  centrifuged  the  mixture.  To  the 
separated  clear  fluid  they  again  added  some  rab- 
bit red  cells  as  well  as  normal  horse  serum  to 
reactivate  the  mixture.  Horse  serum  is  not  hae- 
molytic for  rabbit  red  cells.  The  mixture  re- 
mained unchanged,  no  haemolysis  taking  place. 
If,  however,  they  added  some  of  this  normal  horse 
serum  to  the  centrifuged  red  cells,  the  latter  imme- 


H&MOLYSINS.  33 

diately  dissolved.  Now,  to  the  clear  centrifuged 
fluid,  which  as  we  have  seen  would  not  dissolve 
rabbit  red  cells,  they  added  guinea-pig  red  cells 
and  again  some  normal  horse  serum  to  reactivate 
the  mixture.  The  guinea-pig  red  cells  all  dis- 
solved. This  proved  conclusively  that  in  the  nor- 
mal goat  serum  there  had  existed  two  specific  inter- 
bodies.  One,  for  rabbit  red  cells,  had  been  tied 
by  these  cells  and  carried  down  with  them  in  cen- 
trifuging;  the  other,  specific  for  guinea-pig  red 
cells,  had  remained  behind. 

Multiplicity  of  the  Active  Substances. — These  in- 
vestigators were  able  to  prove  still  more  in  regard 
to  the  multiplicity  of  the  substances  in  normal 
serum  which  are  concerned  in  haemolysis.  They 
showed  that  beside  the  two  inter-bodies  just  men- 
tioned there  existed  in  goat  serum  two  specific 
complements,  one  for  each  inter-body,  arid  they 
were  able  by  means  of  Pukall  filters  to  separate 
these  two.  In  this  filtration  the  complement  fit- 
ting the  inter-body  for  rabbit  blood  remained 
behind  for  the  greater  part,  while  that  fitting 
the  inter -body  for  guinea-pig  blood  mostly  passed 
through. 

Whereas  then,  according  to  Buchner,  only  one 
substance,  the  alexin,  is  concerned  in  the  haemo- 
lytic  action  of  this  normal  goat  serum,  these  experi- 
ments of  Ehrlich  and  Morgenroth  show  us  four 
substances,  viz.,  two  inter-bodies  and  two  comple- 
ments, This  at  once  makes  clear  the  opposing 


34  IMMUNE  SERA. 

views  of  these  authorities.  But  the  number  of 
active  substances  in  normal  serum  is  still  greater, 
for  in  the  experiments  of  the  last-named  authors 
it  oftens  happens  that  a  specific  inter-body  shows 
itself  to  be  made  up  of  several  inter-bodies,  all,  to 
be  sure,  fitting  the  same  specific  red  cell,  but  dif- 
fering from  each  other  by  their  behavior  toward 
different  complements.  Ehrlich,  therefore,  re- 
gards the  substances  concerned  in  haemolysis 
which  occur  in  normal  serum  to  be  of  great 
number  and  variety.  Buchner  and  Bordet,  on  the 
other  hand,  assume  that  only  one  substance  is 
concerned. 

The  facts  which  we  have  thus  far  developed  in 
regard  to  the  haemolysins  of  normal  serum  apply 
equally  well  to  its  h&magglutinin$.  As  we  men- 
tioned in  the  beginning  of  this  article,  Bordet 
showed  that  not  only  was  the  haemolytic  action  of 
a  specific  serum  increased  for  certain  red  cells,  but 
its  agglutinating  power  was  increased  for  the  same 
cell.  According  to  this,  then,  as  a  result  of  the 
immunizing  process  there  are  formed  not  only 
haemolysins  but  also  haemagglutinins. 

Haemagglutinins  of  Normal  Serum. — Analogous  to 
the  haemolytic  action  of  normal  serum  on  the  red 
cells  of  certain  other  species,  we  find  that  nor- 
mal serum  is  able  to  agglutinate  the  red  cells  of 
many  other  species  and  bacteria.  For  example, 
normal  goat  serum  agglutinates  the  red  cells  of 
man,  pigeon,  and  rabbit;  normal  rabbit  serum 


HMMOLYSINS.  35 

agglutinates  typhoid  and  cholera  bacilli.  Bordet 
could  show  that  the  bacterial  agglutinins  are  gov- 
erned by  the  same  laws  of  combination  that  Ehr- 
lich  and  Morgenroth  showed  governed  the  inter- 
bodies  of  normal  sera.  Thus  if  to  a  normal  serum 
which  agglutinates  both  typhoid  and  cholera  ba- 
cilli some  typhoid  bacilli  be  added  and  the  mixture 
centrifuged,  the  clear  fluid  will  no  longer  be  able 
to  agglutinate  typhoid  bacilli.  It  will  still,  however, 
readily  agglutinate  those  of  cholera.  The  typhoid 
agglutinin  has  in  this  way  been  tied  to  the  typhoid 
bacilli  first  added,  and  with  them  it  has  been  car- 
ried down  in  the  centrifuged  sediment.  If  the 
experiment  be  reversed,  so  that  cholera  bacilli  are 
first  added  and  then  the  mixture  centrifuged,  the 
clear  fluid  will  contain  the  typhoid  agglutinin,  but 
not  that  of  cholera.  These  points,  brought  out  by 
Bordet  for  bacterial  agglutinins,  I  have  had  Mal- 
koff  study  regarding  the  haemagglutinins  of  nor- 
mal serum,  and  this  investigator  has  found  the 
same  facts  to  apply  to  these  substances.  To  nor- 
mal goat  serum,  which  agglutinates  the  red  cells 
of  man,  rabbits,  and  pigeons,  he  added  human  red 
cells  and  then  centrifuged  the  mixture.  In  this 
way  the  agglutinin  for  these  cells  was  abstracted 
from  the  serum,  which  then  was  capable  of  agglu- 
tinating the  red  cells  of  rabbits  and  pigeons,  but 
incapable  of  agglutinating  human  red  cells.  When 
he  used  pigeon  blood  instead  of  the  human  blood, 
the  agglutinin  for  pigeon  blood  was  abstracted, 


36  IMMUNE  SERA. 

leaving  the  agglutinins  for  the  red  cells  of  man 
and  rabbits,  etc. 

These  experiments  of  Bordet  and  Malkoff  on 
the  selective  combination  of  the  cells  show  that 
with  the  agglutinins  as  with  the  lysins  (solvent 
substances)  it  is  a  question  of  numerous  substances 
and  not  of  a  single  one.  When,  for  example,  nor- 
mal goat  serum  is  able  simultaneously  to  aggluti- 
nate several,  say  three,  species  of  blood-cells,  this 
action  is  not  due  to  a  single  agglutinin  which  af- 
fects all  three  species,  but  is  the  work  of  three  dis- 
tinct substances,  each  specific  for  a  certain  red 
cell. 

Nature  of  the  Agglutinins. — The  agglutinins  are 
fairly  resistant  substances  which  withstand  heat- 
ing to  60°  C.,  and  lose  their  power  only  on  heating 
to  65°  C.  It  is  possible,  therefore,  to  make  a  serum 
haemolytically  inactive  by  heating  to  55°  C.,  and 
still  preserve  its  agglutinating  power.  Corre- 
sponding to  the  specific  combining  power  of  these 
agglutinins,  they  possess  a  haptophore  group 
which  effects  the  combination,  and  a  second  group, 
easily  decomposed  by  acids,  which  effects  the 
clumping.  In  the  bacterium  as  well  as  in  the 
blood-cell  there  exists  a  substance  not  yet  closely 
studied,  called  the  agglutinable  substance.  This 
also  has  two  groups,  a  haptophore,  which  com- 
bines with  the  haptophore  group  of  the  agglutinin; 
and  a  second,  more  delicate  group,  which  is  acted 
on  by  the  functional  group  of  the  agglutinin. 


H&MOLYSINS.  37 

These  points  have  only  very  recently  bean  brought 
out  by  Eisenberg  and  Kraus,  and  by  the  author. 

This  agglutination  then  is  a  chemical  combina- 
tion between  the  agglutinating  substance  of  the 
serum  and  the  agglutinable  substance  of  the  red 
cell  or  bacterium  and  it  proceeds  in  definite  chem- 
ical proportions.  The  chemical  and  physical  as- 
pects of  the  process  itself  are  still  the  subject  of 
various  theories.  These  I  shall  not  discuss  here, 
as  they  lack  experimental  support.  The  relation 
of  the  agglutinins  to  the  precipitins  is  still  ob- 
scure, so  that  I  shall  not  venture  an  opinion  on  the 
subject. 

Agglutinoids. — Agglutinins  which  have  lost  their 
agglutinophore  group  through  the  action  of  acids, 
etc.,  but  which  still  possess  their  haptophore 
group,  are  called  agglntinoids,  just  as  toxins  which 
have  lost  their  toxophore  group  are  called  toxoids. 
Such  agglutinoids,  then,  may  still  combine  with 
the  blood-cells  or  bacteria  without,  however,  being 
able  to  produce  any  clumping  or  agglutination. 

Purpose  of  Agglutination.  —  It  is  not  yet  clear 
what  the  purpose  of  the  agglutinating  function  is. 
Gruber,  the  first  to  thoroughly  study  and  appreci- 
ate the  bacterial  agglutinins,  assumes  that  the 
process  injures  the  affected  cell,  preparing  it  for 
solution  and  destruction.  After  numerous  experi- 
ments I  have  not  been  able  to  convince  myself  of 
any  damaging  influence  of  the  agglutinins  on  the 
affected  cell,  be  this  blood-cell  or  bacterium,  and 


3  IMMUNE  SERA. 

the  observations  of  other  authors  confirm  this  opin- 
ion. Agglutinated  bacteria  are  capable  of  living 
and  of  reproduction,  and  agglutinated  red  blood- 
cells  are  no  more  fragile  or  easier  to  destroy  than 
normal,  not  agglutinated  cells.  Neither  can  any- 
thing be  discovered  microscopically  which  would 
indicate  any  injury  to  their  structure. 

One  thing  is  certain:  that  the  agglutinins  are  in 
no  way  related  to  the  lysins  found  in  serum, 
and  so,  of  course,  are  not  identical  with  these.  The 
simultaneous  occurrence  in  a  serum  of  immune 
bodies,  inter-bodies,  complements,  and  agglutinins 
is  an  entirely  independent  phenomenon  which  is  no 
way  regular.  There  are  sera  which  dissolve  certain 
cells  without  agglutinating  them,  and  others  which 
agglutinate  cells  without  dissolving  them. 

Difference  between  a  Normal  and  a  Specific  Immune 
Serum.  —  Practical  Application.  —  Returning  now  to 
the  question  of  the  difference  between  a  specific 
immune  serum  and  a  normal  one,  we  find  this  to  be 
as  follows:  Normal  serum  contains  a  great  variety 
of  inter-bodies,  in  very  small  amounts,  and  a  consid- 
erable amount  of  complements.  In  immune  serum, 
on  the  other  hand,  the  amount  of  a  specific  inter- 
body,  the  one  which  fits  the  haptophore  group  of  a 
certain  cell,  is  enormously  increased.  This  spe- 
cifically increased  inter-body,  it  will  be  remembered, 
is  called  the  immune  body.  The  complement,  as 
shown  by  v.  Dungern,  Bordet,  Ehrlich  and  Morgen- 
roth,  and  myself,  is  in  no  way  increased  by  the  im- 


39 

munizing  process.     The""iiic Jea^eT  affects  solely  the 
immune  body.     It  is  therefore  possible  to  have  a 
serum   which   contains   more   immune   body   than 
complement  to  satisfy  it,  and  if  we  withdraw  such  a 
serum  from  an  animal  we  shall  find  that  it  contains 
some  free  immune  body.     This  serum  can  only  then 
exert  its  full  power  when  the  full  amount  of  comple- 
ment is  present,  i.e.,  when  some  normal  serum  is 
added.     If  we  treat  a  rabbit  with  the  red  cells  of  an 
ox,  as  v.  Dungern  did,  we  shall  obtain  a  serum  which 
is  haemolytic  for  ox  blood.     0.05  c.c.  of  this  freshly 
drawn  serum  suffices  to  dissolve  5.0   c.c.   of  a  5% 
mixture  of  ox  blood.     If  now  we  add  to  this  haemo- 
lytic serum  a  little  normal  rabbit  serum,  we  shall 
find  that  only  one-tenth  of  the  amount  of  serum  is 
required;    i.e.,  only  0.005  c-c-  to  dissolve  the  same 
quantity  of  ox  blood.     This  means  that  through 
the  addition  of  the  rabbit  serum,  which,  of  course, 
is  not  haemolytic  for  ox  blood,  a  sufficient  amount 
of  complement  was  added  to  enable  all  the  immune 
body  of  the  specific  serum  to  act.     This  specifically 
increased  power  of  the  immune  serum  to  act  on 
certain  definite  cells  depends  on  the  fact  that  the 
immune    body    resulting     from    the    immunizing 
process   concentrates    the    action    of    the    comple- 
ment   scattered   through   the    serum,  on    cells   for 
which  it  has  definite  affinities.     If  2  c.c.  of  normal 
guinea-pig  serum  are  able  to  dissolve,  we  will  say, 
5  c.c.  of  a  5%  defibrinated  rabbit-blood  mixture, 
and  if  we  find  that  after  the  immunizing  process 


40  IMMUNE  SERA. 

0.05  c.c.  of  the  guinea-pig  serum  suffice  to  dissolve 
the  same,  amount  of  rabbit  blood,  we  conclude 
that  through  this  process  the  inter-body,  i.e.  the 
immune  body,  has  been  increased  forty  times.  We 
know  that  the  complement  has  not  been  increased, 
but  this  is  now  able  to  act  by  means  of  forty  times 
increased  combining  facilities.  This  increase,  how- 
ever, is  exclusively  for  rabbit  blood-cells.  In  a 
bactericidal  immune  serum  this  specific  increase  is 
sometimes  as  much  as  100,000  times  that  of  normal 
serum.  The  practical  idea  to  be  gained  from  this 
for  the  therapy  of  infectious  diseases  is  this:  that 
with  the  injection  of  an  immune  serum  we  supply 
only  one  of  the  necessary  constituents  to  kill  and 
dissolve  the  bacteria,  and  that  is  the  immune  body. 
We  do  not,  however,  supply  the  second,  i.e.  the 
complement,  for  this  we  have  seen  is  not  increased 
by  the  immunizing  process.  As  matters  stand, 
then,  the  use  of  a  specified  immune  serum  for  thera- 
peutic purposes  assumes  that  the  complement 
which  fits  exactly  to  the  immune  body  and  which  is 
essential  for  the  latter's  action  will  be  found  in  the 
organism  to  be  treated.  Because  in  certain  infec- 
tious diseases  the  required  complement  is  present 
in  too  small  amounts  in  the  organism,  I  have  sug- 
gested that  the  curative  power  of  many  bacteri- 
cidal sera  might  be  increased  by  the  simultaneous 
injection  of  the  sera  of  certain  normal  animals  in 
order  to  gain  in  this  way  an  increased  amount  of 
complement ;  but  we  shall  soon  see  that  this  pro- 


B&MOLYSINS.  41 

cedure,  while  of  great  value  in  animal  experiments, 
presents  certain  difficulties. 

All  that  has  here  been  said  regarding  the  specific 
increased  haemolytic  power  of  sera  applies  equally 
to  the  specific  increased  agglutinating  power  follow- 
ing the  injection  of  animals  with  certain  cells.  As 
a  result  of  such  injections,  that  agglutinin  which 
stands  in  specific  relation  to  the  blood-cell  injected 
is  increased  according  to  the  laws  of  the  side-chain 
theory,  and  such  a  serum  therefore  possesses  an 
increased  agglutinating  power  for  these  particular 
cells.  With  the  agglutinins  this  increase  in  power 
is  sometimes  an  enormous  one.  If,  for  example,  a 
normal  serum  is  just  able  to  agglutinate  a  certain 
cell  when  diluted  i  to  10  with  normal  salt  solution, 
it  is  possible  by  means  of  the  immunizing  process  to 
obtain  a  serum  which  in  dilutions  of  one  to  several 
thousands  will  still  completely  agglutinate  the  cells. 

In  such  a  sketch  as  this,  I  cannot  dwell  on 
the  practical  importance  that  this  specific  increased 
agglutinating  power  has  in  the  serum  diagnosis  of 
certain  infectious  diseases,  such  as  typhoid,  etc. 

Nature  of  the  Immune  Body — Partial  Immune  Bodies 
of  Ehrlich. — Turning  now  to  a  closer  study  of  the 
nature  of  the  immune  body,  we  again  find  a  differ- 
ence of  opinion.  Whereas  Bordet,  Metchnikoff, 
and  Besredka  assume  each  immune  body  to  be  a 
single  definite  substance,  Ehrlich  and  Morgenroth 
as  a  result  of  their  experiments  hold  to  a  plurality 
of  bodies.  These  authors  say  that  each  immune 


42  IMMUNE  SERA. 

body  is  built  up  of  a  number  of  partial-immune 
bodies,  a  point  to  which  we  have  already  alluded. 
In  support  of  this  view  they  offer  the  following  ex- 
periment. On  immunizing  a  rabbit  with  ox  blood, 
they  obtained  a  serum  hasmolytic  not  only  for  ox 
blood  but  also  for  goat  blood;  on  immunizing  a 
rabbit  with  goat  blood  they  obtained  a  serum  hasmo- 
lytic for  goat  blood  and  ox  blood.* 

According  to  Ehrlich's  theory,  then,  the  red  cells 
of  the  ox  possess  certain  receptors  which  are  identi- 
cal with  receptors  possessed  by  the  goat  red  cells. 
From  this  it  follows  that  in  a  single  red  cell  there 
are  several  or  many  groups  each  of  which  is  able, 
when  it  finds  a  fitting  receptor,  to  take  hold  of  a 
single  immune  body.  Ehrlich  and  Morgenroth, 
therefore,  claim  that  the  immune  body  of  a  haemo- 
lytic  serum  is  composed  of  the  sum  of  the  partial 
immune  bodies  which  correspond  to  the  individual 
receptors  used  to  excite  the  immunity.  It  may  be 
assumed,  then,  that  not  all  of  the  combining  groups 
of  a  cell,  be  this  a  blood-cell  or  a  bacterium,  will 
find  fitting  receptors  in  every  animal  organism, 
and  that  therefore  not  all  the  possible  partial  im- 
mune bodies  will  be  equally  developed.  In  one 
animal  there  may  be  receptors  which  are  not  pres- 
ent in  another,  and  in  this  way  there  might  be  a  dif- 
ferent variety  of  partial  immune  bodies  in  the  two 


*  We  have  already  called  attention  to  these  exceptions 
to  the  rule  of  specific  action. 


HMMOLYSINS.  43 

animals.  This  would  lead  to  the  possibility  of  the 
occurrence  of  immune  bodies,  for  the  same  species 
of  blood-cell  or  bacterium,  differing  from  each  other 
in  the  partial  immune  bodies  composing  them, 
according  to  the  variety  of  animals  used  in  prepar- 
ing the  serum. 

MetchnikofFs  Views — Practical  Importance  of  the 
Point. — This  view  is  directly  opposed  to  that  of 
Metchnikoff  and  Besredka,  who  believe  that  a  cer- 
tain immune  body,  e.g.  one  specific  for  ox  blood, 
is  always  the  same  no  matter  from  what  animal  it 
is  derived.  The  point  is  not  merely  theoretical, 
but  under  certain  circumstances  of  great  practical 
importance.  If  we  believe,  as  Ehrlich  does,  that 
the  immune  body  differs  according  to  the  species  of 
animal  from  which  it  is  derived,  i.e.,  that  it  is  made 
up  of  different  partial-immune  bodies,  then  we  must 
admit  that  we  have  better  chances  for  finding  fit- 
ting complements  if  we  make  use  of  immune  bodies 
derived  from  a  variety  of  animals.  We  would,  for 
instance,  be  likely  to  achieve  better  results  in  treat- 
ing a  typhoid  patient  with  a  mixture  of  specific 
bactericidal  typhoid  sera  derived  from  a  variety  of 
animals  than  if  we  used  a  serum  derived  only  from 
a  horse.  For  in  such  a  mixture  of  immune  bodies 
the  variety  of  partial-immune  bodies  must  be  very 
great  and  the  chances  that  the  complements  of  the 
human  body  will  find  fitting  immune  bodies,  and  so 
lead  to  the  destruction  of  the  typhoid  bacilli,  are 
greatly  increased.  Ehrlich  and  his  pupils  have 


44  IMMUNE  SERA. 

actually  proposed  such  a  procedure  in  the  use  of 
bactericidal  sera  for  therapeutic  purposes.* 

Support  for  Ehrlich's  View. — Besides  the  above 
experiments  we  possess  others  which  support  the 
theory  that  the  immune  body  is  not  a  simple  but 
a  compound  substance,  v.  Dungern  had  already 
shown  that  following  the  treatment  of  an  animal 
with  ciliated  epithelium  from  the  trachea  of  an  ox, 
there  were  developed  immune  bodies  which  acted 
not  only  on  the  ciliated  epithelium  but  also  on  the 
red  cells  of  oxen.  We  must  assume,  therefore,  that 
the  ciliated  epithelium  and  the  red  cells  of  the  ox 
possess  common  receptors.  Analogous  to  this  is 
the  action  of  the  immune  body  resulting  from  the 
injection  of  spermatozoa,  as  was  pointed  out  by 
Metchnikoff  and  Moxter. 

We  see,  then,  that  the  specific  action  of  immune 
bodies  is  not  so  limited  as  to  apply  only  to  the  cells 
used  in  the  immunizing  process,  but  extends  to 
other  cells  which  have  receptors  in  common  with 
these,  t 

*  Reasoning  along  similar  lines,  namely,  that  the  human 
complement  must  fit  the  immune  body  of  the  therapeutic 
serum,  Ehrlich  has  also  proposed  that  these  bactericidal 
sera  be  derived  from  animals  very  closely  related  to  man, 
e.g.,  apes,  etc. 

t  The  same  holds  good  for  the  agglutinins  and  the  pre- 
cipitins  still  to  be  studied.  In  these  the  action  extends 
also  to  closely  related  cells  and  bacteria,  or  in  the  case  of 
the  precipitins  to  closely  related  albumins,  as  these  possess 
a  number  of  receptors  which  are  common  to  them  and  to 
the  cells  or  substances  used  for  immunizing. 


H&MOLYSINS.  45 

Coming  now  to  the  question  as  to  what  part  of 
the  cell  it  is  which  excites  the  production  of  the 
haemolytic  immune  body,  we  find  this,  according  to 
v.  Dungern,  to  be  the  stroma  of  the  red  cells.  If 
this  be  so,  it  must  be  the  stroma  which  combines 
with  the  immune  body.  Nolf,  however,  claims 
that  the  cell  contents  are  factors  in  the  production 
of  the  immune  body.  So  far  as  concerns  the  site 
in  the  organism  where  the  substances  used  in  immu- 
nizing find  their  receptors,  this  is  not  known  for 
the  haemolytic  immune  body.  For  the  bacteri- 
cidal immune  bodies  of  cholera  and  typhoid,  how- 
ever, we  know  from  the  researches  of  Pfeiffer,  Marx, 
and  myself  that  this  is  chiefly  in  the  bone  marrow 
as  well  as  in  the  spleen  and  lymph  bodies. 

Anti-haemolysins :  their  Nature — Anti-complement  or 
Anti-immune  Body? — A  further  step  in  the  study 
of  haemolysins  is  one  discovered  independently 
by  Ehrlich  and  Morgenroth  on  the  one  hand  and 
Bordet  on  the  other.  These  authors  succeeded  in 
producing  an  anti-H&molysin.  The  procedure  is 
closely  related  to  the  results  gained  by  immunization 
against  bacterial  poisons.  A  specific  haemolysin, 
one,  for  example,  specific  for  rabbit  blood,  de- 
rived by  treating  a  guinea-pig  with  rabbit  red 
cells,  is  highly  toxic  to  rabbits.  Injected  into  the 
animals  intravenously  in  doses  of  5  c.c.  it  kills  the 
animals  acutely,  causing  intra  vitam  a  solution  of 
the  red  cells.  Such  a  haemolytic  serum,  then,  acts 
the  same  as  a  bacterial  poison,  and  it  is  possible  to 


4$  IMMUNE  SERA. 

immunize  against  this  just  as  well  against  a  bac- 
terial poison.  For  example,  to  keep  to  our  illustra- 
tion, rabbits  are  injected  first  with  very  small  doses 
of  this  specific  haemolytic  serum.  The  dose  is 
gradually  increased  until  it  is  found  that  the  animal 
tolerates  amounts  that  would  be  absolutely  fatal  to 
animals  not  so  treated.  If  some  of  the  serum  of 
this  animal  is  now  abstracted  and  added  to  the 
specific  hsemolytic  serum,  it  is  found  that  the  power 
of  the  latter  will  be  inhibited.  This  shows  that  an 
anti-h&molysin  has  been  formed.  As  we  know  that 
the  action  of  the  haemolysin  depends  on  the  com- 
bined action  of  two  substances,  the  immune  body 
and  the  complement,  the  question  arises  to  which 
of  these  two  the  anti -haemolysin  is  related.  Is  it 
an  anti -immune  body  or  an  an  ti  -complement  ?  A 
study  of  this  question  has  shown  that  both  these 
substances  are  present.  In  the  serum  of  the  rabbit 
treated  with  specific  haemolysin,  both  an  anti- 
immune  body  and  an  anti-complement  have  been 
found.  For  the  details  of  the  experiments  of  Ehr- 
lich  and  Morgenroth  and  of  Besredka,  which  dem- 
onstrated this,  I  must  refer  to  the  original  articles. 
The  first-named  authors  were  further  able  to  show 
that  the  action  of  the  anti -complement  depended 
on  a  haptophore  group  which  it  possessed,  enabling 
it  to  combine  with  the  haptophore  group  of  the 
complement,  thus  satisfying  this  and  hindering  its 
combination  with  the  complementophile  group  of 
the  immune  body  (see  figure). 


H&MOLYSINS.  47 

Anti-complement.  —  Since  the  complements  are 
constituents  of  normal  serum,  it  should  be  possible 
to  produce  anti-complements  by  injecting  animals 
merely  with  normal  serum ;  and  they  can,  in  fact, 
be  so  produced.  If  rabbits  are  treated  by  inject- 
ing them  several  times  with  normal  guinea-pig 
serum,  a  serum  may  be  obtained  from  these  rabbits 
which  contains  anti-complements  against  the  com- 
plements of  normal  guinea-pig  serum.  A  serum 

n. 
[ 

COMPLEMENT 

COMPLEMENT 

ANTICOMPLEMENT 
IMMUNE    BODY 


CELL  m  m      {•:•  U  CELL 

FlG.   3.     (After  Levaditi.) 

obtained  in  this  way  of  course  contains  only  one  of 
the  anti-haemolytic  bodies,  the  anti-complement, 
and  not  the  anti-immune  body.  This  is  because 
normal  serum  is  too  poor  in  immune  body  (inter- 
body)  to  excite  the  production  of  any  anti-immune 
body. 

If  to  a  haemolytic  serum  derived  from  guinea-pigs 
we  add  an  anti-complement  serum  derived,  as  just 
stated,  from  rabbits  and  containing  an  anti-com- 
plement specific  for  guinea-pig  complement,  the 


4§  IMMUNE  SERA. 

haemolytic  action  of  the  former  will  be  inhibited  for 
the  reason  that  the  complement  necessary  for  the 
haemolysis  to  take  place  has  been  bound  by  the 
anti-complement.  (See  Fig.  3.)  One  must,  how- 
ever, observe  the  precaution  to  heat  the  anti-com- 
plement serum  of  the  rabbit  to  55°  C.  before  so  mix- 
ing it,  in  order  to  destroy  the  complement  which  it 
contains  and  which  would  otherwise  reactivate  the 
guinea-pig  immune  body. 

From  the  foregoing  we  see  that  either  anti- 
immune  body  alone  or  anti -complement  alone  is 
able  to  inhibit  the  haemolytic  action.  Haemolysis 
cannot  take  place  when  either  of  the  two  necessary 
factors  is  bound  and  prevented  from  acting.* 

The  anti-complements  are  specific  bodies,  i.e.,  an 
anti-complement  combines  only  with  its  specific 
complement.  Thus  an  anti-complement  serum 
derived  from  rabbits  by  treatment  with  guinea- 
pig  serum  combines  only  with  the  complement  of 
normal  guinea-pig  serum,  not,  however,  with  the 
complements  of  other  animals.  Exceptions  to  this 
are  those  cases  in  which  the  complement  of  the 
other  species  possess  receptors  identical  with  those 
of  the  first. 

*  By  treating  animals  with  normal  sera  of  certain  other 
species  it  is  possible  to  produce  not  only  anti-complements 
but  also  specific  anti-bodies  against  certain  other  con- 
stituents of  normal  serum.  These  are,  for  example,  anti- 
agglutinins,  which  inhibit  the  action  of  the  hasmagglu- 
tinins  of  normal  serum,  and  anti-precipitins,  which  we  shall 
discuss  later. 


H&MOLYSINS.  49 

In  order  that  a  normal  serum  of  species  A,  in- 
jected into  species  B,  produce  anti-complements 
there,  the  side-chain  theory  demands  that  the  com- 
plements of  A  find  fitting  receptors  in  species  B. 
According  to  Ehrlich,  however,  normal  serum  con- 
tains many  different  complements  and  not  merely  a 
single  one.  Under  the  circumstances  it  is  easily 
possible  that  only  a  few  of  the  complements  in  the 
serum  of  A  find  fitting  receptors  in  species  B.  We 
shall  then  obtain  an  anti-complement  serum  which 
inhibits  the  action  of  some  but  not  of  all  the  comple- 
ments of  species  A.  Thus  it  might  inhibit  the 
action  of  a  complement  fitting  to  a  certain  bacteri- 
cidal immune  body,  and  not  of  one  contained  in  the 
same  serum  which  fitted  a  certain  haemolytic  im- 
mune body,  etc. 

Auto-anticomplements.  --  A  question  of  great 
practical  importance  now  arises.  Is  it  possible 
under  certain  conditions  for  an  organism  to  manu- 
facture within  itself  anti-complements  against  its 
own  complements,  i.e.,  auto-anticomplements?  The 
complements,  owing  to  their  ferment-like  digestive 
power,  must  play  an  important  role  in  the  living 
organism ;  for  this  concerns  itself  not  only  with  the 
destruction  of  bacteria,  etc.,  an  important  factor  in 
the  natural  immunity  against  diseases,  but  also, 
according  to  Ehrlich,  Buchner,  and  the  author,  with 
the  solution  and  digestion  of  all  kinds  of  foreign 
albuminous  bodies  which  enter  the  organism.  Any 
inhibition  of  this  important  function  would  there- 


50  IMMUNE  SERA. 

fore  be  followed  by  severe  disturbances,  particu- 
larly, however,  by  a  decreased  resistance  against 
infectious  diseases.  The  author  succeeded  in  dem- 
onstrating that  animals  injected  with  anti-comple- 
ments to  tie  up  their  complements  were  much  less 
resistant  to  certain  infectious  diseases. 

The  spontaneous  development  in  an  animal  of 
auto-anticomplement,  i.e.,  substances  developed 
within  the  organism  against  its  own  complements, 
has  not  yet  been  demonstrated.  Ehrlich  and  Mor- 
genroth  were  able  in  a  rabbit  to  excite  the  produc- 
tion of  an  auto-anticomplement  by  treating  the 
animal  in  a  certain  way.  Ordinarily,  normal  rab- 
bit serum  is  slightly  solvent  for  guinea-pig  blood. 
If  the  rabbits  are  treated  with  goat  serum,  the  rab- 
bit serum  loses  this  solvent  power  for  guinea-pig 
red  cells.  Even  if  fresh  normal  rabbit  serum  is  now 
added  the  hsemolysis  does  not  take  place,  although 
we  know  that  this  fresh  serum  is  haemolytic.  This 
shows  that  in  the  serum  of  the  rabbit  treated  with 
goat  blood  an  anti-complement  has  been  formed 
which  combines  with  the  complement  of  normal 
rabbit  blood,  for  it  was  able  to  inhibit  the  action 
of  the  complement  of  the  normal  freshly  added 
rabbit  serum.  In  the  rabbit's  body,  then,  as  a  result 
of  this  procedure,  an  anti-complement  has  been 
formed  against  the  complement  of  its  own  serum,  a 
true  auto-anticomplement. 

Now,   according  to  the  side-chain  theory,   there 
are  no  receptors  in  an  organism  for  the  complements 


H&MOLYSINS.  51 

of  the  same  organism.  The  formation  of  these 
auto-anticomplements,  according  to  Ehrlich,  can 
only  be  explained  by  assuming  that  in  normal  goat 
serum  there  are  present  complements  which  are 
almost  identical  with  those  of  the  rabbit  serum, 
but  which  differ  from  them  in  that  they  find  recep- 
tors in  the  rabbit  serum  whose  haptophore  group 
fits  to  their  own. 

Fluctuations  in  the  Amount  of  the  Active  Substances 
in  Serum. — As  already  said,  we  have  thus  far  been 
unable  to  show  that  the  complements  of  an  organ- 
ism are  decreased  through  the  action  of  spontane- 
ously formed  anti-complements.  We  have,  how- 
ever, come  to  know  certain  conditions  under  which 
there  may  be  a  decrease  of  certain  complements  in 
normal  serum.  Ehrlich  and  Morgenroth  showed 
that  in  rabbits  poisoned  with  phosphorus  and  in 
whom,  therefore,  the  liver  was  badly  damaged,  the 
serum  on  the  second  day  (the  height  of  the  disease) 
had  lost  its  power  to  dissolve  guinea-pig  blood, 
and  that  this  was  due  to  a  disappearance  of  the 
complement.  Metchnikoff  also  reported  that  in  an 
animal  suffering  from  a  continuing  suppurating 
process  the  complement  had  fallen  considerably  in 
amount.  Especially  interesting  are  the  experi- 
ments of  v.  Dungern,  who  showed  that  animal  cells, 
hence  emulsions  of  fresh  organs,  are  able  to  attract 
and  combine  with  complements. 

Fully  as  important  as  the  question  of  a  decrease 
in  complements  or  an  inhibition  of  their  action,  is 


52  IMMUNE  SERA. 

that  of  the  possibility  to  artificially  increase  them. 
A  number  of  authors,  among  them  Nolf  and  Muller, 
have  answered  this  question  in  the  affirmative. 
They  believe  they  have  noticed  such  an  increase 
following  the  injection  of  an  animal  with  all  sorts 
cf  substances,  such  as  normal  serum  of  another 
animal,  sterile  bouillon,  etc.  v.  Dungern,  as  well 
as  myself  and  others,  have  not  been  able  to  convince 
ourselves  of  the  possibility  of  such  a  definite  in- 
crease. I  tried  to  excite  the  increased  production 
of  complement  by  injecting  guinea-pigs  for  some 
time  with  anti-complement.  This  being  the  oppo- 
site of  the  complement,  I  hoped  to  be  able  by  im- 
munizing to  excite  an  increase  of  the  complements. 
In  this  I  was  unsuccessful,  though  of  course  it  may 
be  possible  with  another  species  of  animal. 

Despite  all  this  we  must  believe  that  the  amount 
of  complement  as  well  as  the  amount  of  other  active 
substances  of  the  blood,  inter-bodies,  agglutinins, 
anti -toxins,  ferments,  anti-ferments,  etc.,  is  subject 
to  great  fluctuations  even  in  the  same  individual, 
a  constant  change  going  on  within  the  organism. 
Ehrlich,  in  particular,  has  pointed  out  these  indi- 
vidual and  periodic  variations  and  has  insisted  on 
their  importance.  Very  likely,  under  circumstances 
of  which  we  now  know  very  little,  these  substances 
are  at  certain  times  produced  in  greater  amounts, 
at  other  times  in  lesser;  sometimes  they  may  be 
absent  entirely  in  an  individual  in  whom  they  were 
previously  present.  For  example,  the  serum  of  a 


HMMOLYSINS.  53 

dog  will  at  times  dissolve  the  red  cells  of  cats,  rab- 
bits, and  guinea-pigs,  at  other  times  not.  Further- 
more, the  serum  of  one  and  the  same  animal  may 
possess  specific  haemolytic  properties  for  certain 
cells,  and  later  on  may  lose  this  property  entirely. 
In  human  serum  these  same  individual  and  periodic 
variations  may  be  demonstrated,  as  I  was  able  by 
many  experiments  to  prove.  However,  the  cir- 
cumstances on  which  these  variations  depend  are  as 
yet  entirely  unknown  to  us.  Possibly  we  are  deal- 
ing here  with  subtle  pathological  changes. 

Source  of  the  Complements — Leucocytes  as  a  Source — 
Other  Sources. — Where  do  the  complements  or  alexins 
originate?  This  question  has  been  studied  particu- 
larly by  Metchnikoff  and  by  Buchner,  also  by  Bail, 
Hahn,  Schattenfroh,  and  others.  These  investiga- 
tors believe  that  the  leucocytes  are  the  source  of 
the  complements  or  alexins.  There  is,  however, 
this  difference  between  the  views  of  MetchnikofI 
and  Buchner ;  whereas  Buchner  believes  the  alexins 
to  be  true  secretory  products,  Metchnikoff  believes 
that  they  originate  on  the  breaking  up  of  the  leuco- 
cytes, i.e.,  that  they  are  decomposition  products. 
Metchnikoff  bases  his  belief  chiefly  on  the  work  of 
his  pupil,  Gengou,  who  showed  that  although  the 
serum  was  rich  in  alexin  (i.e.,  complement)  the 
plasma  contained  none  at  all. 

Other  authors,  as  Pfeiffer  and  Moxter,  as  a  result 
of  their  experiments,  are  not  willing  to  assume  the 
existence  of  any  relationship  between  the  alexins 


54  IMMUNE  SERA. 

and  the  leucocytes.  Gruber  as  well  as  Schatten- 
froh  are  ready  to  believe  the  leucocytes  to  be  the 
source  of  an  alexm,  but  claim  that  this  is  different 
from  that  found  in  serum.  I  myself  believe  that 
the  leucocytes  are  a  source  of  complements  (alexins) ; 
for  I  succeeded  in  producing  anti-complement  by 
means  of  injections  of  pure  leucocytes  which  had 
been  washed  free  from  all  traces  of  serum,  and 
which  had  been  obtained  by  injections  of  aleu- 
ronat.  Because  of  the  plurality  of  the  complements, 
I  have  expressed  the  view  that  the  leucocytes  are 
probably  one  source,  but  not  the  only  one,  for  the 
complements  of  the  serum.  Landsteiner  and 
Donath  have  confirmed  my  views  experimentally. 
They  succeeded  in  producing  anti-complement  by 
the  injection  not  only  of  leucocytes,  but  of  other 
animal  cells.  Furthermore,  the  experiments  of 
Ehrlich  and  Morgenroth  already  mentioned,  in 
which  the  complements  disappeared  after  the  de- 
struction of  the  liver  function,  show  that  the  liver 
cells  are  concerned  in  the  formation  of  complements. 
Structure  of  Complements — Haptophore  and  Zymo- 
toxic  Groups  —  Complementoids.  —  The  structure  of 
the  complement  has  been  studied  particularly  by 
Ehrlich  and  Morgenroth,  and  by  P.  Miiller.  We 
have  seen  that  the  complements  lose  their  power 
when  heated  to  55°  C.  If,  however,  we  inject  ani- 
mals with  a  normal  serum  that  has  previously  been 
heated  to  55°  C.,  we  shall  still  excite  in  these  ani- 
mals the  production  of  anti-complements.  This 


H&MOLYS1NS.  55 

shows  that  the  heating  has  not  destroyed  the  entire 
complement  body,  but  only  that  part  which  effects 
the  digesting,  solvent  action.  The  part  of  the 
complement  concerned  with  the  combination  with 
the  inter-body  or  immune  body,  in  other  words, 
that  part  called  by  Ehrlich  the  haptophore  group, 
must  have  remained  intact.  It  is  clear,  therefore, 

zymotoxic  group 
COMPLEMENT 
haptophore  group 

IMMUNE   BODY 
FIG.  4. 

that  anti-complements  can  only  be  formed  when 
there  remain  in  the  complements  haptophore  groups 
that  fit  certain  receptors  in  the  organism  of  the 
animal  injected.  From  this  it  follows  that  the 
complements  consist  of  a  combining  haptophore 
group  which  withstands  heating  to  55°  C.,  and  an- 
other more  fragile  group  which  possesses  the  actual 
solvent  properties,  and  which  Ehrlich  calls  the 
zymotoxic  group.  There  is  a  perfect  analogy  be- 
tween this  and  the  toxins  already  studied.  These, 
it  will  be  remembered,  consist  of  a  haptophore  and 


5  6  IMMUNE  SERA. 

a  toxophore  group.  And  just  as  those  toxins  which 
had  lost  their  toxophore  group  were  called  toxoids, 
so  Ehrlich  and  Morgenroth  purpose  to  call  comple- 
ments which  have  lost  their  zymotoxic  group  com- 
plementoids. 

Isolysins  —  Autolysins  —  Anti-isolysins. —  All  of  the 
preceding  studies  in  haemolysis  have  concerned 
themselves  with  the  results  obtained  by  injecting 
animals  of  one  species  with  blood-cells  of  another. 
Ehrlich  and  Morgenroth  now  sought  to  discover 
what  the  result  would  be  if  they  injected  an  animal 
with  blood-cells  of  its  own  species.  They  injected 
goats  with  goat  blood,  and  found  that  when  the 
amount  injected  at  one  time  was  large  the  serum 
of  the  goat  injected  acquired  haemolytic  properties 
for  the  blood  of  many  other  goats  but  not  for  all. 
The  substances  thus  formed  the  authors  called 
isolysins.  These,  then,  are  substances  which  will 
dissolve  the  blood  of  other  individuals  of  the  same 
species.  Substances  which  dissolve  the  blood-cells 
of  the  same  individual  are  called  autolysins.  But 
autolysins  have  so  far  been  demonstrated  experi- 
mentally only  once  (by  Ehrlich  and  Morgenroth). 
If  one  tests  the  properties  of  an  isolysin  of  a  goat  on 
the  blood  of  a  great  many  other  goats,  it  will  be 
found  that  this  will  be  strongly  solvent  for  the 
blood  of  some,  slightly  for  the  blood  of  others,  and 
not  at  all  for  still  others. 

By  using  a  blood  that  was  readily  dissolved  by 
the  isolysin,  and  proceeding  in  the  same  series  of 


HMMOLYSINS.  57 

experiments  which  we  have  already  studied  under 
haemolysis,  Ehrlich  and  Morgenroth  showed  that 
the  isolysins,  like  the  hsemolysins,  consist  of  an 
immune  body  and  a  complement  of  the  normal 
serum.  The  experiments  undertaken  by  these 
authors  were  made  on  thirteen  goats  and  the  sur- 
prising fact  developed  that  the  thirteen  resulting 
isolysins  were  all  different.  For  example,  the  iso- 
haemolytic  serum  of  one  goat  dissolved  the  red  cells 
of  goats  A  and  B;  that  of  a  second  goat  those  of 
C  and  D ;  of  a  third  those  of  A  and  D,  but  not  of  C, 
and  so  on.  If  now  they  produced  anti -isolysins  by 
injecting  animals  with  these  isolysins,  they  found 
that  these  anti-isolysins  were  specific;  i.e.,  the  anti- 
isolysin  of  A  would  inhibit  the  action  only  of  iso- 
lysin  of  A,  but  not  of  C,  etc.  These  results  are  of 
the  highest  clinical  interest,  for  they  show  a  differ- 
ence in  similar  cells  of  the  same  species,  something 
that  had  never  before  been  suspected.  In  the 
above  the  blood -cells  of  species  A  must  have  a  dif- 
ferent biological  constitution  than  those  of  species 
C,  etc. 

The  fact  that  after  injections  of  large  amounts  of 
cells  of  the  same  species  isolysins  develop,  but  that 
autolysins  are  almost  never  formed,  caused  Ehr- 
lich and  Morgenroth  to  assume  that  the  body  pos- 
sesses distinct  regulating  functions  which  naturally 
prevent  the  formation  of  the  highly  destructive 
autolytic  substance.  It  is  obvious  that  if  there 
were  no  such  regulating  facilities  the  absorption  of 


58  IMMUNE  SERA. 

large  bloody  effusions  and  haemorrhages  might  lead 
to  the  formation  by  the  organism  of  autolysins 
against  its  own  blood-cells.  Gengou,  a  pupil  of 
Metchnikoff,  in  a  very  recent  work,  believes  to  have 
shown  experimentally  that  the  destructive  action 
of  these  autolysins  is  hindered  by  the  simultaneous 
production  of  an  auto-anti-immune  body  which 
immediately  inhibits  their  action. 

In  order  that  isolysins  may  be  formed,  it  seems 
necessary  to  overwhelm  the  organism  once  or  sev- 
eral times  with  large  amounts  of  cells  or  cell  prod- 
ucts of  the  same  species;  to  produce,  as  Ehrlich 
says,  an  ictus  imniunisatorius .  I  tried,  by  using 
various  blood  poisons,  such  as  haemolytic  sera, 
toluylenediamine,  etc.,  for  a  continued  length  of 
time,  to  cause  the  formation  of  these  isolysins,  but 
without  success,  although  in  these  experiments 
each  injection  was  followed  by  an  appreciable  de- 
struction of  red  cells  and  absorption  of  their  decom- 
position products.  The  gradual  and  even  repeated 
absorption  of  not  too  large  quantities  of  decom- 
posed red  cells  does  not  therefore  lead  to  the  forma- 
tion of  isolysins;  but,  as  already  said,  a  sudden 
overwhelming  of  the  organism  by  large  amounts 
of  the  cells  or  their  products  is  necessary. 


II.  CYTOTOXINS. 

Cytotoxins — Definition — Leucotoxin — Nature  of  the 
Cytotoxin — Anti-cytotoxin. — After  it  had  been  found 
that  the  injection  of  an  animal  with  red  blood- 
cells  of  another  animal  was  followed  by  the  produc- 
tion of  definite,  specific  reaction  substances,  investi- 
gators experimented  to  see  whether  this  was  also 
the  case  if  other  animal  cells  were  used.  Injections 
were  made  with  white  blood-cells,  spermatozoa  of 
other  animals,  etc.,  and  there  resulted  a  series  of 
reaction  substances,  entirely  analogous  to  the  hag- 
molysins,  which  were  specific  for  the  cells  used  for 
injection.  These  sera  Metchnikoff  calls  cytotoxins. 
After  Delezenne  had  published  a  short  article  on  a 
serum  haemolytic  for  white  blood-cells,  Metchnikoff 
undertook  a  study  of  the  substances  produced  in 
sera  of  animals  treated  with  leucocytes  of  another 
species.  He  injected  guinea-pigs  with  the  mesen- 
teric  glands  and  bone  marrow  of  a  rabbit.  He 
also  injected  for  several  weeks  half  an  Aselli's  pan- 
creas at  a  time,  at  intervals  of  four  days.  If  he 
withdrew  serum  from  such  a  guinea-pig  he  found 
this  to  be  intensely  solvent  for  white  blood-cells  of 
a  rabbit.  He  called  this  serum  leucotoxin.  This 

59 


60  IMMUNE  SERA. 

leucotoxin  is  very  poisonous  for  these  animals,  and 
kills  them  within  a  few  hours.  Non-fatal  doses  at 
first  excite  a  marked  hypoleucocytosis,  which  is 
followed  after  a  few  days  by  a  compensatory  hyper- 
leucocytosis.  Leucotoxin  destroys  the  mononu- 
clear  as  well  as  the  polynuclear  leucocytes  of  the 
animal,  as  was  shown  by  Funk.  Leucotoxin  which 
had  been  derived  by  injection  of  the  leucocytes  of 
horses,  oxen,  sheep,  goats,  or  dogs  acted  only  on 
the  leucocytes  of  that  species,  not  on  the  leucocytes 
of  man.  So  far  as  the  mechanism  of  the  cytotoxic 
action  is  concerned,  it  has  been  found  that  this  is 
the  same  as  that  of  the  haemolysins.  The  action  of 
the  specific  cytotoxic  serum  is  always  due  to  the 
combined  action  of  two  substances  in  the  serum,  a 
specific  immune  body,  and  an  alexin  or  comple- 
ment present  also  in  normal  serum.  The  cyto- 
toxic sera,  like  the  haemolytic  sera,  are  rendered 
inactive  by  heating  to  55°  C.  In  other  respects 
also  the  cytotoxic  sera  maintain  the  analogy  to 
the  haemolytic  sera.  Thus  it  is  possible  by  immu- 
nizing with  a  cytotoxin  to  obtain  an  anti-cyto- 
toxin.  MetchnikofI,  for  example,  was  able  to  pro- 
duce an  anti-leucotoxin  by  injecting  animals  -with 
leucotoxin.  This  anti-body  inhibited  the  action  of 
the  leucotoxin. 

Spermotoxin. — Another  specific  cell-dissolving  se- 
rum was  produced  by  Landsteiner,  Metchnikoff, 
and  Moxter,  by  injecting  animals  with  the  sperma- 
tozoa of  other  animals.  Such  a  serum  rapidly 


CYTOTOXINS.  6 1 

destroys  the  spermatozoa  of  the  animals  whose 
product  was  injected.  This  cytotoxin  was  named 
spermotoxin.  If  animals  are  treated  with  spermato- 
zoa there  is  produced  a  serum  which  is  not  only  a 
spermotoxin,  but  which  is  also  haemolytic  for  the 
red  cells  of  that  animal.  This  was  demonstrated 
by  Metchnikoff  and  Moxter,  and  has  already  been 
referred  to  in  discussing  haemolysins.  If,  for  ex- 
ample, we  inject  the  spermatozoa  of  sheep  into 
rabbits,  we  shall  obtain  a  serum  that  is  spermo- 
toxic  for  sheep,  as  well  as  haemolytic  for  sheep 
red  cells.  This  is  the  case  even  when  the  greatest 
care  is  exercised  to  exclude  every  trace  of  blood 
in  obtaining  and  injecting  the  spermatozoa.  The 
haemolysin,  however,  differs  somewhat  from  that 
obtained  by  injecting  sheep  red  cells,  and  its  pro- 
duction is  not  hard  to  explain  if  we  hold  fast  to  the 
side-chain  theory.  We  assume  that  these  sperma- 
tozoa possess  certain  receptors  in  common  with  the 
red  blood-cells  of  the  same  animal. 

Anti-spermotoxin  —  Auto-spermotoxin. — By  treating 
an  animal  with  its  spermotoxin  we  can  produce 
an  anti-spermotoxin  which  will  inhibit  the  action 
of  the  former.  Metalnikoff,  a  pupil  of  Metchnikoff, 
has  demonstrated  the  occurrence  of  auto-spermo- 
toxin.  This,  however,  is  only  of  scientific  interest, 
and  I  mention  it  here  only  for  the  sake  of  com- 
pleteness. 

Cytotoxin  for  Epithelium. — v.  Dungern  has  pro- 
duced an  anti-epithelial  serum  by  treating  animals 


62  IMMUNE  SERA. 

with  the  ciliated  tracheal  epithelium  of  oxen.  This 
serum  is  rapidly  destructive  for  this  particular  kind 
of  epithelium.  In  this  serum  there  is  formed  at 
the  same  time  a  specific  haemolytic  body,  just  as  m 
the  case  of  the  spermotoxic  serum,  and  for  the  same 
reasons.  The  ciliated  epithelium  possesses  a  recep- 
tor group  common  to  it  and  to  the  red  blood-cells. 
It  is  therefore  able  to  produce  an  immune  body 
which  is  made  up  of  two  partial  immune  bodies. 

This  anti-epithelial  serum  is  of  further  interest  in 
that  it  leads  us  to  hope  that  eventually  we  shall  be 
able  to  produce  sera  which  are  cytotoxic  for  other 
varieties  of  epithelial  cells,  particularly  those  of 
pathological  origin,  as  carcinoma.  There  have,  how- 
ever, been  no  further  contributions  to  this  subject 
worthy  of  mention. 

Lindemann,  a  pupil  of  Metchnikoff,  and  Nefe- 
dieff  have  treated  animals  with  an  emulsion  of 
kidney  cells  of  another  species,  and  have  found 
that  a  serum  was  produced  which  was  specific 
against  the  kidney  cells  of  the  second  animal,  and 
which  produced  an  albuminuria  in  these  animals 
intra  vitam. 

In  similar  fashion,  Delezenne  and  Deutsch  have 
produced  a  serum  specifically  cytotoxic  for  liver 
cells. 

Neurotoxin. — Delezenne  and  Madame  Metchni- 
koff have  injected  animals  with  central-nervous- 
system  substance,  and  so  produced  a  specific  neuro- 
toxin.  They  injected  clucks  intraperitoneally,  giv- 


CYTOTOXINS.  63 

ing  them  five  or  six  injections  of  10  to  20  grammes 
of  dog  brain  and  spinal  cord  mixed  with  normal 
salt  solution.  The  serum  of  these  ducks  injected 
intracerebrally  into  dogs  in  doses  of  one-half  c.c. 
caused  the  dogs  to  die  almost  at  once  in  complete 
paralysis,  whereas  if  normal  duck  serum  was  in- 
jected in  the  same  way  no  effects  of  any  kind  were 
produced.  If  smaller  doses  of  the  specific  neuro- 
toxic  serum  were  administered,  say  o.i  to  0.2  c.c., 
various  paralyses  and  epileptiform  convulsions  set 
in,  from  which  the  animals  sometimes  recovered. 
The  action  of  this  serum  is  also  specific,  i.e.,  the 
serum  of  ducks  treated  with  dog  brain  causes 
these  symptoms  only  in  dogs,  while  on  rabbits  it 
acts  no  differently  than  normal  duck  serum. 

These  are  the  most  important  of  the  cytolytic 
or  cytotoxic  sera,  though,  of  course,  the  list  can 
readily  be  extended  experimentally.  In  all  this 
we  are  evidently  dealing  with  a  general  biological 
law  which  we  can  express  somewhat  as  follows: 
An  animal,  species  A,  into  whose  body  are  injected 
cells  or  cell  products  of  species  B,  reacts  by  producing 
specific  substances  in  its  serum  against  these  cells  or 
cell  products,  provided,  of  course,  that  the  incorpo- 
rated cells  or  cell  products  find  fitting  receptors  in  the 
body  of  A. 

Practical  Applications  of  the  Cytotoxins. — With  a 
number  of  these  sera  therapeutic  experiments 
have  been  made  in  the  human  subject  Those  of 
Metchnikoff  and  his  pupil  Besredka  deserve  men- 


64  IMMUNE  SERA. 

tion.  They  used  a  haemolytic  serum  derived  from 
goats  which  had  been  treated  with  human  blood. 
One  volume  of  this  was  able  to  dissolve  an  equal 
volume  of  human  blood  within  a  few  minutes.  This 
specific  serum  was  injected  into  patients  suffering 
from  lepra,  in  doses  of  one-half  to  seven  c.c.,  sub- 
cutaneously.  As  a  result  there  was,  of  course,  first 
a  reduction  of  red  cells,  due  to  haemolysis.  This  was 
followed  after  about  six  days  by  an  increase  of  the 
same,  and  also  by  an  increase  in  their  haemoglobin 
content.  Certain  other  symptoms  which  showed 
themselves  on  the  leprous  nodules  Metchnikoff 
ascribes  to  the  leucotoxin  which  the  serum  contains. 
These  experiments  have,  however,  had  no  further 
practical  results. 


III.  PRECIPITINS. 

Definition. — All  of  the  foregoing  experiments  have 
concerned  themselves  with  the  results  obtained  by 
injection  of  cellular  material  of  one  animal  into 
another.  In  the  further  study  of  this  subject,  ex- 
periments were  made  to  discover  what  happens 
when  dissolved  albuminous  bodies  of  one  species  are 
injected  into  animals  of  another  species.  This  line 
of  investigation  was  first  pursued  by  Tsistowitsch,  <^~ 
who  injected  rabbits  with  ,the  serum  of  horses  and 
of  eels.  On  withdrawing  serum  from  such  rabbits 
and  mixing  it  with  horse  or  eel  serum,  the  mix- 
ture became  cloudy,  owing  to  the  precipitation  of  L 
part  of  the  albumin  of  the  horse  or  eel  serum  by 
that  of  the  rabbit.  Normal  rabbit  serum  does  not 
possess  this  property.  Bordet  was  able  to  demon- 
strate that  the  same  thing  takes  place  if  rabbits  are 
treated  with  chicken  blood.  On  mixing  such  a 
serum  with  chicken  serum  a  precipitate  formed. 
These  substances  which  develop  in  the  serum  by 
treating  an  animal  with  albuminous  bodies  of  an- 
other animal,  and  which  precipitate  these  albumins  ^ 
when  the  sera  of  the  two  animals  are  mixed,  are 

65 


66  IMMUNE  SERA. 

called  precipitins*  This  power  of  the  organism  to 
react  to  the  injection  of  foreign  dissolved  albumin- 
ous substances  has  been  found  to  be  very  extensive. 
Lactoserum  —  Other  Specific  Precipitins.  —  Bordet, 
by  injecting  cows'  milk  into  rabbits,  was  able  to 
produce  a  serum  which  precipitates  the  casein  of 
cows'  milk.  He  called  this  lactoserum.  Ehrlich, 
Morgenroth,  the  author,  Schutze,  Myers,  and  Uhlen- 
huth  showed  that  by  treating  a  rabbit  with  chicken 
albumin  a  precipitin  is  formed  which  precipi- 
tates chicken  albumin.  Myers,  by  treating  animals 
with  Witte's  pepton  and  globulin,  produced  a  serum 
that  contained  specific  anti-peptons  and  anti- 
globulins.  Pick  and  Spiro,  by  using  albumose, 
produced  anti-albumoses.  Lechainche  and  Vallee, 
Stern,  Mertens,  and  Zulzer  treated  animals  with 
human  albuminous  urine  and  produced  a  serum 
which  contained  a  precipitin  specific  for  this  sub- 
stance. Schutze,  by  treating  rabbits  with  a  vege- 
table albumin,  as  well  as  with  human  myoalbumin, 
produced  a  precipitin  specific  for  these  albumins. 
This  does  not  exhaust  the  recital  of  the  work  done 
in  this  field,  and  there  is  a  host  of  other  albuminous 
bodies  which,  when  injected  into  an  animal,  are 
able  to  excite  the  production  of  specific  precipitins. 
The  production  of  precipitins  for  the  albuminous 

*  It  will  be  recalled  that,  besides  the  production  of  pre- 
cipitins, the  above  procedure  causes  the  formation  of  other 
anti-bodies  such  as  anti-complements,  anti-agglutinins, 
etc. 


PRECIPITINS.  67 

bodies  found  in  bacterial  cultures  had  previously 
been  shown  by  Kraus. 

Nature  of  the  Precipitins. — The  precipitins  are 
fairly  resistant  bodies,  whose  power  gradually  de- 
clines at  a  temperature  of  60°  C.,  but  is  not  lost  until 
70°  C.  is  reached.  The  resulting  precipitate  is  solu- 
ble in  weak  acids  and  alkalies.  Peptic  digestion 
destroys  the  substances  which  effect  the  precipita- 
tion. Concerning  the  chemical  nature  of  the  pre- 
cipitins, we  have  an  admirable  study  by  Leblanc, 
who  finds  in  the  case  of  a  large  number  of  precipi- 
tins that  they  are  precipitated  with  that  fraction  of  £^ 
the  serum  which  Hofmeister  calls  the  pseudo-globu- 
lins. The  pseudo-globulins  constitute  that  part  of 
the  total  globulins  which  is  soluble  in  distilled 
water,  while  the  rest,  the  euglobulins,  are  insoluble. 
The  nature  of  the  resulting  precipitate  has  also  been 
studied  by  Leblanc.  He  finds  that  it  is  a  combina- 
tion of  the  precipitated  albumin  with  the  anti-body, 
a  pseudo-globulin,  of  the  specific  serum.  In  this 
combination  the  properties  of  the  pseudo-globulin 
predominate,  showing  that  it  is  the  specific  serum  ^ 
which  furnishes  the  greater  part  of  the  precipitate. 

Action  Not  Entirely  Specific. — Of  special  interest 
is  the  inquiry  as  to  how  far  the  action  of  these  pre- 
cipitins is  specific.  The  first  experiments  of  Bordet 
had  shown  that  the  specificity  is  not  complete ;  that 
the  serum  of  rabbits  injected  with  chicken  serum 
is  a  precipitin  not  only  for  chicken  serum  but 
also  for  that  of  pigeons.  The  author  and  Schutze, 


68  IMMUNE  SERA. 

as  well  as  Stern,  were  able  to  show  that  the  serum 
of  rabbits  treated  with  guinea-pig  serum  is  a 
preeipitin  also  for  the  serum  of  monkeys.  Prob- 
ably this  is  because  the  sera  of  guinea-pigs  and  of 
monkeys  possess  common  receptors.  For  this  rea- 
son also  the  injection  of  a  serum  from  one  animal 
into  a  closely  related  animal  does  not  excite  the 
production  of  a  preeipitin,  e.g.,  the  serum  of  chickens 
injected  into  pigeons  or  that  of  rabbits  into  guinea- 
pigs.  We  may  assume  that  in  these  closely  related 
animals  the  serum  of  the  one  fails  to  find  receptor 
groups  with  which  to  combine;  on  the  contrary,  it 
finds  groups  similar  to  its  own,  and  these  cannot 
react  on  one  another.  In  line  with  this  is  the  work 
of  Uhlenhuth,  who  showed  that  the  serum  of  ani- 
mals treated  with  chicken  egg  albumin  is  a  pre- 
eipitin for  egg  albumin  of  other  closely  related 
birds. 

Practical  Application. — These  precipitins  have  very 
recently  found  a  practical  application.  Fish,  Ehr- 
lich,  Morgenroth,  the  author,  and  Schiitze  inves- 
tigated the  specific  action  of  lactoserum.  They 
found  that  a  serum  derived  by  treating  an  animal 
with  cows'  milk  contained  a  preeipitin  which  reacted 
only  on  the  casein  of  cows'  milk,  but  not  on  that  of 
human  milk  or  goats'  milk.  The  serum  of  an  ani- 
mal treated  with  human  milk  was  specific  for  the 
casein  of  human  milk,  etc.  Ehrlich,  Morgenroth, 
and  the  author  also  experimented  with  the  serum 
resulting  from  treatment  with  chicken  egg  albumin, 


PRECIPi  TINS.  69 

and  found  that  this,  while  not  strictly  specific  so  far 
as  closely  related  species  are  concerned,  is  yet  so 
against  other  species.  The  precipitins,  therefore, 
react  on  closely  related  albumins,  but  are  absolutely 
specific  against  those  of  unrelated  species. 

Wassermann's  Method  of  Differentiating  Albumins — 
To  Test  Blood  Stains. — As  a  result  of  these  re- 
searches the  author  proposed ,.  at  the  Congress  for 
Internal  Medicine,  1900,  to  use  these  sera  as  a  means 
of  differentiating  albumins,  i.e.,  to  distinguish  the 
different  albumins  from  one  another  and  particu- 
larly to  distinguish  those  derived  from  man  from 
those  of  other  animals.  This  proposal  thus  to  use 
the  Tsistowitsch-Bordet  precipitins  had  important 
practical  and  theoretical  results.  Uhlenhuth,  Was- 
sermann,  Schutze,  Stern,  Dieudonne,  and  others 
showed  that  a  serum  could  be  produced  from  rab- 
bits by  injecting  them  with  human  serum,  by  means 
of  which  it  is  possible  to  tell  positively  whether  a 
given  old,  dried  blood  stain  is  human  blood  or  not. 
The  procedure  is  as  follows:  The  suspected  clot  is 
mixed  with  a  small  quantity  of  normal  salt  solution 
and  then  filtered.  To  some  of  this  in  a  test-tube 
about  the  double  the  amount  of  the  specific  serum 
(derived  as  above)  is  added.  The  specific  serum  is 
first,  however,  tested  as  to  its  activity.  As  a  con- 
trol test  we  place  a  little  blood  of  another  species, 
e.g.,  of  an  ox,  in  a  second  test-tube  together  with 
some  of  the  specific  serum  and  a  little  normal  salt 
solution.  In  a  third  test-tube  we  place  some  of  the 


7°  IMMUNE  SERA. 

suspected  blood  solution,  and  in  a  fourth  some  of 
the  specific  serum,  both  without  any  additions. 
All  four  tubes  are  placed  in  an  incubator  at  37°  C. 
for  one  hour  or  are  left  at  normal  room  temperature 
for  several  hours.  If  the  clot  be  that  of  human 
blood,  the  first  tube  must  show  distinct  evidences 
of  precipitation,  while  all  the  control  tubes  must 
have  remained  clear.  This  reaction  is  absolutely 
specific,  with  one  exception,  as  was  pointed  out  by 
the  author,  Schutze,  and  Stern.  A  serum  derived 
by  treating  an  animal  with  human  serum  reacts 
also  to  the  serum  of  monkeys.  It  does  not,  how- 
ever, react  to  the  sera  of  any  other  animals  thus 
far  investigated.  This  method,  then,  furnishes 
the  surest  differential  diagnosis  for  forensic  pur- 
poses, as  has  been  proved  by  the  researches  into  the 
subject  by  Ziemke. 

The  Method  Applied  to  Distinguish  Albumins.— 
The  A.  Wassermann  method  of  diagnosis  by  means 
of  precipitins  has  found  further  application.  Le- 
blanc  showed  that  the  serum  of  an  animal  treated 
with  pseudo-globulin  causes  a  precipitate  only  in  a 
solution  of  pseudo-globulin;  one  derived  from  an 
animal  treated  with  serum  albumin,  only  in  a  solu- 
tion of  serum  albumin;  one  derived  by  treatment 
with  haemoglobin,  only  in  a  solution  of  haemoglobin, 
etc.  One  is  enabled,  therefore,  by  this  method  to 
distinguish  the  different  albuminous  bodies.  Le- 
clainche  and  Vallee  as  well  as  Mertens  showed  by 
this  method  that  the  albumins  of  blood  and  milk 


IPRECIPITINS.  71 

are  different,  and  that  therefore  the  albumin  of 
milk  is  not  a  mere  transudation  product,  but  is  a 
true  secretion.  Kowarski  and  Schiitze,  as  already 
mentioned,  could  show  a  difference  between  vege- 
table and  animal  albumin.  Jess  as  well  as  Uhlen- 
muth  used  the  method  to  differentiate  various  kinds 
of  meat  in  the  markets. 

The  principle  and  the  method  are  the  same  in  all 
these  various  applications.  We  treat  animals  with 
the  albumins  which  we  wish  to  differentiate,  and  so 
obtain  sera  specific,  each  for  its  particular  kind  of 
albumin.  These  sera,  then,  produce  precipitates 
only  in  solutions  of  their  respective  albumins.  For 
example,  if  we  wish  to  determine  whether  a  given 
sample  of  meat  is  horse-flesh  or  not  we  must  inject 
an  animal  with  horse  serum,  or,  if  we  prefer,  with 
an  extract  of  horse-flesh.  The  serum  derived  from 
this  animal  will  then  produce  a  precipitate  in  the 
aqueous  extract  of  the  meat  if  this  be  horse-flesh, 
but  not  if  it  be  beef.  Animals  treated  with  dog 
serum  yield  a  serum  which  precipitates  an  aqueous 
extract  of  dog-flesh,  etc.  The  future  will  undoubt- 
edly show  further  practical  applications  of  this 
method. 

Anti-precipitins  —  Iso-precipitins. — Biologically,  the 
precipitins  are  found  to  behave-  like  the  sub- 
stances already  studied.  It  is  possible,  for  exam- 
ple, by  injecting  an  animal  with  a  precipitin,  say 
lactoserum,  to  obtain  an  anti-precipitin,  an  anti- 
lactoserum,  which  counteracts  or  inhibits  the  ac- 


72  IMMUNE  SERA. 

tion  of  the  precipitin.  This  is  entirely  analogous 
to  the  anti-hsemolysins,  the  anti-spermotoxin,  etc. 
If  rabbits  are  treated  with  rabbit  serum,  a  serum 
is  obtained  which  will,  in  certain  cases,  precipitate 
the  serum  of  other  rabbits.  This  was  done  by 
Schiitze,  and  he  called  this  serum  iso-precipitin. 


IV.  CONCLUSION. 

Clinical  Applications  of  Immune  Sera. — In  closing 
this  general  resume  of  the  subject  it  may  be  well 
to  mention  some  of  the  most  important  work 
done  in  the  application  of  these  discoveries  to  clin- 
ical purposes.  Monaco  and  Panichi  have  shown 
that  in  malaria  the  blood  of  the  patients  very  early 
shows  the  presence  of  iso-agglutinins,  so  that  the 
serum  of  these  patients  is  able  to  agglutinate  the 
red  cells  of  other  persons.  Grunbaum  claims  to 
have  proved  the  same  for  typhoid  and  scarlet  fevers. 
The  most  thorough  work  on  this  subject  has  been 
done  by  Eisenberg,  who  after  examining  a  large 
number  of  cases  of  all  kinds  finds  that  iso-agglu- 
tinins and  isolysins  may  be  developed  in  all  dis- 
eases in  which  there  is  destruuction  of  red  blood-cells 
or  other  cell  material  and  a  consequent  absorption 
of  their  products.  According  to  the  studies  thus 
far  made  we  cannot,  in  man,  ascribe  any  specific 
diagnostic  value  to  the  occurrence  of  isolysins  or 
iso-agglutinins.  Rather  may  they  be  regarded  as 
delicate  indicators  which  show  that  there  has  been 
destruction  and  absorption  of  living  cell  material 
in  those  cases  in  which  they  appear. 

73 


74  IMMUNE  SERA. 

Diagnostic  Value  of  Iso-agglutinins  and  of  Isolysins. — 

As  a  result  of  my  own  observations,  made  on 
patients  in  the  Institute  for  Infectious  Diseases, 
I  attach  no  particular  value  to  the  occurrence  of 
iso-agglulinins  in  the  serum  of  one  patient  for  the 
erythrocytes  of  another.  In  this  procedure,  espe- 
cially in  the  case  of  human  erythrocytes,  there  are 
great  sources  of  error.  The  erythrocytes  of  many 
persons  have  an  inclination  to  agglutinate  on  the 
addition  of  any  human  serum,  so  that  even  the  serum 
of  the  same  person  will  agglutinate  these  red  cells. 
In  my  opinion,  therefore,  it  is  unwise  to  attach 
much  importance  to  the  occurrence  of  iso-agglutinins; 
we  should  rather  look  for  the  occurrence  of  isolysins 
in  the  different  diseases.  In  the  isolysins  the  sources 
of  error  just  mentioned  do  not  obtain. 

E.  Neisser,  Doring,  and  Lacqueer  were  able  to 
show  that  in  cases  of  uraemia  substances  appeared 
in  the  serum  which  acted  very  much  like  auto-anti- 
complements.  The  number  of  the  observed  cases, 
however,  is  too  small  to  draw  definite  conclusions. 

Immunization  against  Other  Substances. — For  com- 
pleteness' sake  it  may  be  mentioned  that  it  has 
been  possible  to  immunize  against  a  great  vari- 
ety of  ferments;  thus  against  emulsin  (Hilde- 
brandt),  certain  ferments  in  bacteria  (v.  Dungern), 
rennet  (Morgenroth  and  Briot),  and  against  fibrin 
ferment  (Bordet  and  Gengou). 

With  all  these  sera  the  results  obtained  experi- 
mentally have  thus  far  been  applied  clinically  to  too 


CONCLUSION.  75 

few  cases  to  allow  of  any  conclusive  opinion  as  to 
the  merits  of  this  method  of  treatment.  Without 
doubt  carefully  conducted  researches  in  this  field 
might  lead  to  discoveries  of  great  value  to  the  sick, 
and  give  us  an  insight  into  most  delicate  disturb- 
ances of  the  organism. 


LITERATURE. 

Belfanti  and  Carbone.     Jorn.  d.  la  R.  Acad.  d.  Med.  di 

Torino,  1898,  No.  8. 
Bordet.     Ann.  de  1'Inst.  Pasteur,  1895,  1898,  1899,  1899, 

1900. 

Bordet  and  Gengou.     Ann.  de  1'Inst.  Pasteur,  1901. 
Buchner.     Verhandlungen     des     Kongresses     fur     Innere 

Medicin,  1892;  Munch,  med.  Wochenschr. ,  1891,  1899. 
Delezenne.     Comptes  rendus  de  TAcad.  des  Sciences,  1900. 
Ehrlich.     Klin.  Jahrbuch,  1898. 
Ehrlich.     Schlussbetrachtungen    in    NOTHNAGEL'S    Patho- 

logie  und  Therapie,  1901. 
Ehrlich.     Croonian  Lecture,  1900. 
Ehrlich  and  Morgenroth.     Berl.  klin.  Wochenschr.,   1899, 

1899,  1900,  1900,  1901,  1901. 
Eisenberg.     Wiener  klin.  Wochenschr.,  1901. 
Fish.     Boston  Journal  of  Medicine,  1900. 
Kraus.     Wiener  klin.  Wochenschr.,  1899. 
Landsteiner.     Centralbl.  f.  Bakteriologie,  1899. 
Leblanc.     La  Cellule,  T.  XVIII,  1901. 
Malkoff.     Deutsche  med.  Wochenschr.,  1900. 
Metchnikoff.     Ann.  de  1'Inst.  Pasteur,   1899,   1900,   1900; 

Revue  Gene"r.  des  Sciences  pures  et  appliquees,  1901. 
Monaco  and  Panichi.     Rendic.  delle  R.  Acad.  dei  Lincei, 

1900. 

Moxter.     Deutsche  med.  Wochenschr.,  1900. 
Muller.     Centralbl.  f.  Bakteriologie,  1901. 

76 


LITERATURE.  77 

Myers.     Centralbl.  f.  Bakteriologie,  1900. 
Nolf.     Ann.  de  1'Inst.  Pasteur,  1900,  1900. 
Tsistowitsch.     Ann.  de  1'Inst.  Pasteur,  1899. 
Uhlenhuth.     Deutsche    med.    Wochenschr.,     1900,     1901, 

1901. 

v.  Dungern.     Munch,  med.  Wochenschr.,  1899,  1899,  1900. 
Wassermann.     Zeitschr.  f.  Hygiene,  Bd.  37;    Kongress  f. 

Innere  Medicin,  1900.     S.  50. 
Wassermann  and  Schutze.     Berl.  klin.  Wochenschr.,  1901. 


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Principles  of  Animal  Nutrition 8vo,  4  oo 

Budd  and  Hansen's  American  Horticultural  Manual: 

Part  I. — Propagation,  Culture,  and  Improvement i  amo,  50 

Part  II. — Systematic  Pomology i2mo,  50 

Downing's  Fruits  and  Fruit-trees  of  America 8vo,  oo 

Elliott's  Engineering  for  Land  Drainage i2mo,  50 

Practical  Farm  Drainage i2mo,  oo 

Green's  Principles  of  American  Forestry I2mo,  50 

Grotenfelt's  Principles  of  Modern  Dairy  Practice.     (Woll.) i2mo,  oo 

Kemp's  Landscape  Gardening 1 2ino,  50 

Maynard's  Landscape  Gardening  as  Applied  to  Home  Decoration 12 mo,  50 

Sanderson's  Insects  Injurious  to  Staple  Crops i2mo,  50 

Insects  Injurious  to  Garden  Crops.     (In  preparation.) 
Insects  Injuring  Fruits.     (In  preparation.) 

Stockbridge's  Rocks  and  Soils 8vo,  50 

WolTs  Handbook  for  Farmers  and  Dairymen i6mo,  50 

ARCHITECTURE. 

Baldwin's  Steam  Heating  for  Buildings 12010,  2  50 

Berg's  Buildings  and  Structures  of  American  Railroads 4to,  5  oo 

Birkmire's  Planning  and  Construction  of  American  Theatres 8vo,  3  oo 

Architectural  Iron  and  Steel 8vo,  3  50 

Compound  Riveted  Girders  as  Applied  in  Buildings 8vo,  2  oo 

Planning  and  Construction  of  High  Office  Buildings 8vo,  3  50 

Skeleton  Construction  in  Buildings 8vo,  3  oo 

Briggs's  Modern  American  School  Buildings 8vo,  oo 

Carpenter's  Heating  and  Ventilating  of  Buildings 8vo,  oo 

Freitag's  Architectural  Engineering.     2d  Edition,  Rewritten 8vo,  50 

Fireproofing  of  Steel  Buildings 8vo,  50 

French  and  Ives's  Stereotomy 8vo,  50 

Gerhard's  Guide  to  Sanitary  House-inspection i6mo,  oo 

Theatre  Fires  and  Panics I2mo .  50 

1 


Holly's  Carpenters'  and  Joiners'  Handbook i8mo,  o  75 

Johnson's  Statics  by  Algebraic  and  Graphic  Methods 8vo'  2  oo 

Kidder's  Architect's  and  Builder's  Pocket-book.     (Rewritten  edition  in  preparation.') 

Merrill's  Stones  for  Building  and  Decoration 8vo,  5  oo 

Monckton's  Stair-building 4to,  4  oo 

Patton's  Practical  Treatise  on  Foundations 8vo,  5  oc 

Siebert  and  Biggin's  Modern  Stone-cutting  and  Masonry 8vo,  i   50 

Snow's  Principal  Species  of  Wood 8vo,  3  50 

Sondericker's  Graphic  Statics  with  Applications  to  Trusses,  Beams,  and  Arches. 

8vo,  2  oo 

Wait's  Engineering  and  Architectural  Jurisprudence 8vo,  6  oo 

Sheep,  6  50 

Law  of  Operations  Preliminary  to  Construction  in  Engineering  and  Archi- 
tecture   8vo,  5  oo 

Sheep,  5  sc 

Law  of  Contracts 8vo,  3  oo 

Woodbury's  Fire  Protection  of  Mills 8vo,  2  50 

Worcester  and  Atkinson's  Small  Hospitals,  Establishment  and  Maintenance, 
Suggestions  for  Hospital  Architecture,  with  Plans  for  a  Small  Hospital. 

i2mo,  i  25 

The  World  VOolumbian  Exposition  offi893 Large  4to,  i  oo 


ARMY  AND  NAVY. 

Bernadou's  Smokeless  Powder,  Nitro-cellulose,  and  theJTheory  of  the  Cellulose 

Molecule i2mo,  2  50 

*  Bruff's  Text-book  Ordnance  and  Gunnery 8vo,  6  oo 

Chase's  Screw  Propellers  and  Marine  Propulsion 8vo,  3  oo 

Craig's  Azimuth 4to,  3  SO 

Cre  lore  and  Squire's  Polarizing  Photo-chronograph 8vo,  3  oo 

Cronkhite's  Gunnery  for  Non-commissioned  Officers 24mol  morocco,  2  oo 

*  Davis's  Elements  of  Law 8vo,  2  50 

*  Treatise  on  the  Military  Law  of  United  States 8vo,  7  oo 

Sheep,  7  50 

De  Brack's  Cavalry  Outpost  Duties.     (Carr.) 24mo  morocco,  2  oo 

Dietz's  Soldier's  First  Aid  Handbook i6mo,  morocco,  i  25 

*  Dredge's  Modern  French  Artillery 4to,.half  morocco,    15  oo 

Durand's  Resistance  and  Propulsion  of  Ships 8vo,  5  oo 

*  Dyer's  Handbook  of  Light  Artillery i2mo,  3  oo 

Eissler's  Modern  High  Explosives 8vo,  4  oo 

*  Fiebeger's  Text-book  on  Field  Fortification Smal^Svo,  2  oo 

Hamilton's  The  Gunner's  Catechism i8mo,  i  oo 

*  Hoff's  Elementary  Naval  Tactics 8vo,  i  50 

Ingalls's  Handbook  of  Problems  in  Direct  Fire 8vo,  4  oo 

*  Ballistic  Tables 8vo,  i  50 

*  Lyons's  Treatise  on  Electromagnetic  Phenomena.   Vols.  I.  and  II.  .8vo,each,  6  oo 

*  Mahan's  Permanent  Fortifications.     (Mercur.) 8vo,  half  morocco,  7  So 

Manual  for  Courts-martial i6mO;.  morocco,  i  50 

*  Mercur's  Attack  of  Fortified  Places I2mo,  2  oo 

*  Elements  of  the  Art  of  War 8vo,  4  oo 

Metcalf 's  Cost  of  Manufactures — And  the  Administration_of  Workshops,  Public 

and  Private 8vo,  5  oo 

*  Ordnance  and  Gunnery i2mo,  5  oo 

Murray's  Infantry  Drill  Regulations i8mo,  paper,  10 

*  Phelps's  Practical  Marine  Surveying 8vo,  2  50 

Powell's  Army  Officer's  Examiner i2mo,  4  oo 

Sl-arpe's  Art  of  Subsisting  Armies  in  War i8mo,  morocco,  i  50 

2 


*  Walke's  Lectures  on  Explosives  .........  .........................  8vo,    4  oo 

*  Wheeler's  Siege  Operations  and  Military  Mining  .....................  8vo, 

Winthrop's  Abridgment  of  Military  Law  ............................  i2mo. 

Woodhull's  Notes  on  Military  Hygiene  .............................  i6mo, 

Young's  Simple  Elements  of  Navigation  ....................  i6mo.  morocco, 


Second  Edition,  Enlarged  and  Revised  ................  i6mo,  morocco, 


ASSAYING. 

Fletcher's  Practical  Instructions  in  Quantitative  Assaying  with  the  Blowpipe. 

1 2mo,  morocco,  I   50 

Furman's  Manual  of  Practical  Assaying 8vo,  3  oo 

Miller's  Manual  of  Assaying i2mo,  i  oo 

O'Driscoll's  Notes  on  the  Treatment  of  Gold  Ores 8vo.  a  oo 

Ricketts  and  Miller's  Notes  on  Assaying 8vo,  3  oo 

Ulke's  Modern  Electrolytic  Copper  Refining 8vo,  3  oo 

Wilson's  Cyanide  Processes 1 2mo,  i  50 

Chlorination  Process i2mo,  i  50 


ASTRONOMY. 

Comstock's  Field  Astronomy  for  Engineers 8vo,  a  50 

Craig's  Azimuth 4to,  3  50 

Doolittle's  Treatise  on  Practical  Astronomy 8vo,  4  oo 

Gore's  Elements  of  Geodesy 8vo,  a  50 

Hayford's  Text-book  of  Geodetic  Astronomy 8vo,  3  oo 

Merriman's  Elements  of  Precise  Surveying  and  Geodesy 8vo,  a  50 

*  Michie  and  Harlow's  Practical  Astronomy 8vo,  3  oo 

*  White's  Elements  of  Theoretical  and  Descriptive  Astronomy xarno.  a  oo 


BOTANY. 

Davenport's  Statistical  Methods,  with  Special  Reference  to  Biological  Variation. 

i6mo,  morocco,  I  35 

Thome  and  Bennett's  Structural  and  Physiological  Botany i6mo,  a  35 

Westermaier's  Compendium  of  General  Botany.     (Schneider.) 8vo,  a  oo 


CHEMISTRY. 

kdriance's  Laboratory  Calculations  and  Specific  Gravity  Tables 12010,  t  35 

Allen's  Tables  for  Iron  Analysis 8vo,  3  oo 

Arnold's  Compendium  of  Chemistry.     (Mandel.)     (In  preparation.') 

Austen's  Notes  for  Chemical  Students i2mo,  i  50 

Bernadou's  Smokeless  Powder. — Nitro-cellulose,  and  Theory  of  the  Cellulose 

Molecule lamo,  2  50 

Bolton's  Quantitative  Analysis 8vo,  i  50 

*  Browning's  Introduction  to  the  Rarer  Elements 8vo,  i  50 

Brush  and  Penfield's  Manual  of  Determinative  Mineralogy 8vo,  4  oo 

Classen's  Quantitative  Chemical  Analysis  by  Electrolysis.  (Boltwood.)  .  . .   8vo,  300 

Cohn's  Indicators  and  Test-papers lamo,  a  oo 

Tests  and  Reagents 8vo,  3  oe 

Copeland's  Manual  of  Bacteriology.     (In  preparation.) 

Craft's  Short  Course  in  Qualitative  Chemical  Analysis.  (Schaeffer.). . .  .i2mo,  i  50 

Drechsel's  Chemical  Reactions.     (Merrill.) I2mo,  i  35 

Duhem's  Thermodynamics  and  Chemistry.     (Burgess.) 8vo,  4  OO 

Eissler's  Modern  High  Explosives 8vo,  4  oo 

3 


Eff rent's  Enzymes  and  their  Applications.     (Prescott.) 8vo,  3  oo 

Erdmann's  Introduction  to  Chemical  Preparations.     (Dunlap.) i2mo,  i   25 

Fletcher's  Practical  InstructionsJn^Quantitative  Assaying  with  the  Blowpipe. 

i2mo,  morocco,  i   50 

Fowler's  Sewage  Works  Analyses lamo,  2  oo 

Fresenius's  Manual  of  Qualitative  Chemical  Analysis.     (Wells.) 8vo,  5  oo 

Manual  of  Qualitative  Chemical  Analysis.     Parti.    Descriptive.     (Wells.) 

8vo,  3  oo 

System   of   Instruction   in    Quantitative    Chemical   Analysis.      (Cohn.) 
2  vols.    (Shortly.) 

Fuertes's  Water  and  Public  Health i2mo,  i  50 

Furman's  Manual  of  Practical  Assaying 8vo,  3  oo 

Gill's  Gas  and  Fuel  Analysis  for  Engineers i2mo,  i   2^5 

Grotenfelt's  Principles  of  Modern  Dairy  Practice.     (Woll.) i2mo.  2  oo 

Hammarsten's  Text-book  of  Physiological  Chemistry.     (Mandel.) 8vo,  4  oo 

Helm's  Principles  of  Mathematical.Chemistry.     (Morgan.) i2mo,  i  50 

Hinds's  Inorganic  Chemistry 8vo,  3  oo 

*  Laboratory  Manual  for  Students I2mo,        75 

Holleman's  Text-book  of  Inorganic  Chemistry.     (Cooper.) 8vo,  2  50 

Text-book  of  Organic  Chemistry.     (Walker  and  Mott.) 8vo,  2  50 

Hopkins's  Oil-chemists'  Handbook 8vo,  3  oo 

Jackson's  Directions  for  Laboratory  Work  in  Physiological  Chemistry .  .8vo,  i  oo 

Keep's  Cast  Iron 8vo,  2  50 

Ladd's  Manual  of  Quantitative  Chemical  Analysis i2mo  i  oo 

Landauer's  Spectrum  Analysis.    (Tingle.) 8vo,  3  oo 

Lassar-Cohn's  Practical  Urinary  Analysis.     (Lorenz.) i2mo.  i  oo 

Leach's  The  Inspection  and  Analysis  of  Food  with  Special  Reference  to  State 

.  Control.     (In  preparation.) 

L»b's  Electrolysis  and  Electrosynthesis  of  Organic  Compounds.  (Lorenz.)  i2mo,  i  oo 

Mandel's  Handbook  for  Bio-cherrical  Laboratory i2mo,  i   so 

*  Martin's  Laboratory  Guide  to  Qualitative  Analysis  with  the  Blowpipe .  .  i2mo,        60 
Mason's  Water-supply.     (Considered  Principally  from  a  Sanitary  Standpoint.) 

3d  Edition,  Rewritten 8vo,  4  oo 

Examination  of  Water.  (Chemical  and  Bacteriological.) i2mo,  i  25 

Meyer's  Determination  of  Radicles^in  Carbon  Compounds.  (Tingle.).  .i2mo,  i  oo 

Miller's  Manual  of  Assaying I2mo,  i  oo 

Mixter's  Elementary  Text-boot1  of  Chemistry i2mo,  i  50 

Morgan's  Outline  of  Theory  of  Solution  and  its  Results 12 mo,  i  oo 

Elements  of  Physical  Chemistry i2mo,  2  oo 

Nichols's  Water-supply,  i  (Considered  mainly  from  a  Chemical  and  Sanitary 

Standpoint,  1883.) 8vo,  2  50 

O'Brine's  Laboratory  Guide'in  Chemical  Analysis 8vo,  2  oo 

O'Driscoll's  Notes  on  the  Treatment  of  Gold  Ores 8vo,  2  oo 

Ost  and  Kolbeck's  Text-book  of  Chemical  Technology.  (Lorenz — Bozart.) 

(In  preparation.) 

*  Penfield's  Notes  on  Determinative  ^Mineralogy  and  Record  of  Mineral  Tests. 

8vo,  paper,        50 

Pictet's   The   Alkaloids  >nd  Ttheir   Chemical   Constitution.      (Biddle.)      (In 
reparation.) 

Pinner's  Introduction  to  Organic  Chemistry.     (Austen.) i2mo,  i  50 

Poole's  Calorific  Power  of  Fuels 8vo.  3  oo 

*  Reisig's  Guide  to  Piece-dyeing 8vo,  25  oo 

Richards  and  Woodman's  Air  .Water,  and  Food  from  a  Sanitary  Standpoint .  8vo,  2  oo 

Richards's  Cost  of  Living  as 'Modified  by  Sanitary  Science i2mo,  i  oo 

Cost  of  Food  a  Study  in  Dietaries I2mo,  i  oo 

*  Richards  and  Williams's  The  Dietary  Computer 8vo,  i  50 

Ricketts'and  Russell's  Skeleton  Notes  upon  Inorganic  Chemistry.     (Part  I. — 

Non-metallic   Elements.) 8vo,  morocco,          75 

4 


Ricketts  and  Miller's  Notes  on  Assaying 8vo,  3  oo 

Rideal's  Sewage  and  the  Bacterial  Purification  of  Sewage 8vo,  ~3  50 

Ruddiman's  Incompatibilities  in  Prescriptions 8vo,  2  oo 

Salkowski's  Physiological  and  Pathological  Chemistry.     (Orndorff.) 

(Shortly.) 
Schimpf  s  Text-book  of  Volumetric  Analysis I2mo, 

Essentials  of  Volumetric  Analysis I2mo, 

Spencer's  Handbook  for  Chemists  of  Beet-sugar  Houses i6mo,  morocco, 

Handbook  for  Sugar  Manufacturers  and  their  Chemists.  .i6mo,  morocco, 
Stockbridge's  Rocks  and  Soils 8vo, 

*  Tillman's  Elementary  Lessons  in  Heat 8vo, 

*  Descriptive  General  Chemistry 8vo  3  oo 

Treadwell's  Qualitative  Analysis.     (HalL) 8vo,  3  oo 

Turneaure  and  Russell's  Public  Water-supplies 8vo,  5  oo 

Van  Deventer's  Physical  Chemistry  for  Beginners.     (Boltwood.) i2mo,  i  50 

*  Walke's  Lectures  on  Explosives 8vo,  4  oo 

Wells's  Laboratory  Guide  in  Qualitative  Chemical  Analysis 8vo,  i  50 

Short  Course  in  Inorganic  Qualitative  Chemical  Analysis  for  Engineering 

Students " i2mo,  i  50 

Whipple's  Microscopy  of  Drinking-water 8vo,  3  50 

Wiechmann's  Sugar  Analysis ; Small  8vo.  2  $u 

Wilson's  Cyanide  Processes izmo,  i  50 

Chlorination  Process i2mo  i  50 

Wulling's  Elementary  Course  in  Inorganic  Pharmaceutical  and  Medical  Chem- 
istry  i amo  2  oo 

CIVIL  ENGINEERING. 

BRIDGES  AND    ROOFS.       HYDRAULICS       MATERIALS   OF   ENGINEERING 
RAILWAY   ENGINEERING. 

Baker's  Engineers'  Surveying  Instruments X2mo,  3  oo 

Bixby's  Graphical  Computing  Table Paper  19^X24!  inches.  25 

**  Burr's  Ancient  and  Modern  Engineering  and  the  Isthmian  Canal.     (Postage , 

27  cents  additional.) 8vo,  net,  3  50 

Comstock's  Field  Astronomy  for  Engineers 8vo,  2  50 

Davis's  Elevation  and  Stadia  Tables 8vo,  i  oo 

Elliott's  Engineering  for  Land  Drainage i  amo,  i  50 

Practical  Farm  Drainage I2mo,  i  oo 

Folwell's  Sewerage.     (Designing  and  Maintenance.) 8vo,  3  oo 

Freitag's  Architectural  Engineering.     2d  Edition,  Rewritten 8vo,  3  50 

French  and  Ives's  Stereotomy 8vo,  2  50 

Goodhue's  Municipal  Improvements I2mo,  I  75 

Goodrich's  Economic  Disposal  of  Towns'  Refuse 8vo,  ,3  50 

Gore's  Elements  of  Geodesy * 8vo,  2  50 

Hayford's  Text-book  of  Geodetic  Astronomy 8vo,  3  oo 

Howe's  Retaining  Walls  for  Earth i2mo,  i  25 

Johnson's  Theory  and  Practice  of  Surveying Small  8vo,  4  oo 

Statics  by  Algebraic  and  Graphic  Methods 8vo,  2  oo 

Kiersted's  Sewage  Disposal I2mo,  i   25 

Laplace's  Philosophical  Essay  on  Probabilities.     (Truscott  and  Emory.)  i2mo,  2  oo 

Mahan's  Treatise  on  Civil  Engineering.     (1873  )     (Wood.) 8vo,  5  oo 

*  Descriptive  Geometry   8vo,  i   50 

Merriman's  Elements  of  Precise  Surveying  and  Geodesy 8vo,  2  50 

Elements  of  Sanitary  Engineering .• 8vo,  2  oo 

Merriman  and  Brooks'*?  Handbook  for  Surveyors i6mo,  morocco,  2  oo 

Nugent's  Plane  Surveying 8vo,  3  50 

Ogden's  Sewer  Design i2mo,  2  60 

Patton's  Treatise  on  Civil  Engineering 8vo  half  leather,  7   50 

5 


Reed's  Topographical  Drawing  and  Sketching 4to,  5  oo 

Rideal's  Sewage  and  the  Bacterial  Purification  of  Sewage 8vo,  3  50 

Siebert  and  Biggin's  Modern  Stone-cutting  and  Masonry 8vo,  i  50 

Smith's  Manual  of  Topographical  Drawing.     (McMillan.) 8vo,  2  50 

Sondericker's   Graphic   Statics,   witn   Applications   to   Trusses.  Beams,   and 

Arches 8vo,  2  oo 

*  Trautwine's  Civil  Engineer's  Pocket-book i6mo,  morocco,  5  oo 

Wait's  Engineering  and  Architectural  Jurisprudence 8vo,  6  oo 

Sheep,  6  50 

Law  of  Operations  Preliminary  to  Construction  in  Engineering  and  Archi- 
tecture  8vo,  5  oo 

Sheep,  5  50 

Law  of  Contracts 8vo,  3  oo 

Warren's  Stereotomy — Problems  in  Stone-cutting 8vo,  2  50 

Webb's  Problems  in  the  U»e  and  Adjustment  of  Engineering  Instruments. 

i6mo,  morocco,  i  25 

*  Wheeler's  Elementary  Course  of  Civil  Engineering 8vo,  4  oo 

Wilson's  Topographic  Surveying 8vo,  3  50 


BRIDGES  AND  ROOFS. 

Boiler's  Practical  Treatise  on  the  Construction  of  Iron  Highway  Bridges.  .8vo,  2  oo 

*         Thames  River  Bridge 4to,  paper,  5  oo 

Burr's  Course  on  the  Stresses  in  Bridges  and  Roof  Trusses,  Arched  Ribs,  and 

Suspension  Bridges .^. . .  8vo,  3  50 

Du  Bois's  Mechanics  of  Engineering.     Vol.  11 Small  4to,  10  oo 

Foster's  Treatise  on  Wooden  Trestle  Bridges 4to,  5  oo 

Fowler's  Coffer-dam  Process  tor  Piers 8vo,  2  50 

Greene's  Roof  Trusses 8vo,  i  25 

Bridge  Trusses 8vo,  2  50 

Arches  in  Wood,  Iron,  and  Stone 8vo,  2  50 

Howe's  Treatise  on  Arches 8vo  4  oo 

Design  of  Simple  Roof-trusses  in  Wood  and  Steel 8vo,  2  oo 

Johnson,  Bryan,  and  Turneaure's  Theory  and  Practice  in  the  Designing  of 

Modern   Framed   Structures Small  4to,  10  oo 

Merriman  and  Jacoby's  Text-book  on  Roofs  and  Bridges: 

Part  I. — Stresses  in  Simple  Trusses 8vo,  2  50 

Part  II. — Graphic  Statics 8vo,  2  50 

Part  III. — Bridge  Design.     4th  Edition,  Rewritten 8vo,  2  50 

Part  IV. — Higher  Structures 8vo,  2  50 

Morison's  Memphis  Bridge 4to,  10  oo 

Waddell's  De  Pontibus,  a  Pocket-book  for  Bridge  Engineers. .  .  i6mo,  morocco,  3  oo 

Specifications  for  Steel  Bridges i2mo,  i  25 

Wood's  Treatise  on  the  Theory  of  the  Construction  of  Bridges  and  Roofs. 8vo,  2  oo 
Wright's  Designing  of  Draw-spans: 

Part  I.  — Plate-girder  Draws 8vo,  2  50 

Part  II. — Riveted-truss  and  Pin-connected  Long-span  Draws 8vo,  2  50 

Two  parts  in  one  volume 8vo,  3  50 


HYDRAULICS. 

Bazm's  Experiments  upon  the  Contraction  of  the  Liquid  Vein  Issuing  from  an 

Orifice.     (Trautwine.) 8vo,  2  oo 

Bovey's  Treatise  on  Hydraulics 8vo,  5  oo 

Church's  Mechanics  of  Engineering 8vo,  6  oo 

Diagrams  of  Mean  Velocity  of  Water  in  Open  Channels paper,  i  50 

6 


Coffin's  Graphical  Solution  of  Hydraulic  Problems i6mo,  morocco,  2  50 

Flather's  Dynamometers,  and  the  Measurement  of  Power i2mo,  3  oo 

Folwell's  Water-supply  Engineering 8vo,  4  oo 

Frizell's  Water-power 8vo,    5  oo 

Fuertes's  Water  and  Public  Health i2mo,  i  50 

Water-filtration  Works iamo,  2  50 

Ganguillet  and  Kutter's  General  Formula  for  the  Uniform  Flow  of  Water  in 

Rivers  and  Other  Channels.     (Hering  and  Trau twine.) 8vo,  4  oo 

Hazen's  Filtration  of  Public  Water-supply 8vo,  3  oo 

Hazlehurst's  Towers  and  Tanks  for  Water- works 8vo,  2  50 

Herschel's  115  Experiments  on  the  Carrying  Capacity  of  Large,  Riveted,  Metal 

Conduits 8vo,  2  oo 

Mason's    Water-supply.     (Considered    Principally   from    a    Sanitary    Stand- 
point.)    3d  Edition,  Rewritten 8vo,  4  oo 

Merriman's  Treatise  on  Hydraulics,     gth  Edition,  Rewritten 8vo,  5  oo 

*  Michie's  Elements  of  Analytical  Mechanics 8vo,  4  oo 

Schuyler's   Reservoirs  for  Irrigation,   Water-power,   and   Domestic   Water- 
supply Large  8vo,  5  oo 

**  Thomas  and  Watt's  Improvement  of  Riyers.     (Post.,  44  c.  additional),  4to,  6  oo 

Turneaure  and  Russell's  Public  Water-supplies 8vo,  5  oo 

Wegmann's  Design  and  Construction  of  Dams 4to,  5  oo 

Water-supply  of  the  City  of  New  York  from  1658  to  1895 4to,  10  oo 

Weisbach's  Hydraulics  and  Hydraulic  Motors.     (Du  Bois.) 8vo,  5  oo 

Wilson's  Manual  of  Irrigation  Engineering Small  8vo,  4  oo 

Wolff's  Windmill  as  a  Prime  Mover 8vo,  3  oo 

Wood's  Turbines 8vo,  a  50 

Elements  of  Analytical  Mechanics 8vo,  3  oo 


MATERIALS  OF  ENGINEERING. 

Baker's  Treatise  on  Masonry  Construction 8vo,  5  oo 

Roads  and  Pavements 8vo,  5  oo 

Black's  United  States  Public  Works Oblong  4to,  5  oo 

Bovev's  Strength  of  Materials  and  Theory  of  Structures 8vo,  7  50 

Burr's  Elasticity  and  Resistance  of  the  Materials  of  Engineering.     6th  Edi- 

tion,  Rewritten 8vo,  7  50 

Byrne's  Highway  Construction 8vo,  5  oo 

Inspection  of  the  Materials  and  Workmanship  Employedln  Construction. 

i6mo,  3  oo 

Church's  Mechanics  of  Engineering ,_ 8vo,  6  oo 

Du  Bois's  Mechanics  of  Engineering.     Vol.  I Small  4to,  7  50 

Johnson's  Materials  of  Construction Large  8vo,  6  oo 

Keep's  Cast  Iron 8vo,  2  50 

Lanza's  Applied  Mechanics 8vo,  7  50 

Martens's  Handbook  on  Testing  Materials.     (Henning.)     2>ols 8vo,  750 

Merrill's  Stones  for  Building  and  Decoration 8vo,  5  oo 

Merriman's  Text-book  on  the  Mechanics  of  Materials 8vo,  4  oo 

Strength  of  Materials I2mo,  i  oo 

Metcalf's  Steel.     A  Manual  for  Steel-users I2mo,  2  oo 

Patton's  Practical  Treatise  on  Foundations 8vo,  5  oo 

Rockwell's  Roads  and  Pavements  in  France i2mo,  i  25 

Smith's  Wire;  Its  Use  and  Manufacture Small  4to,  3  oo 

Materials  of  Machines i2mo,  i  oo 

Snow's  Principal  Species  of  Wood 8vo,  ,j  50 

Spalding's  Hydraulic  Cement i2mo,  2  oo 

Text-book  on^Roads  and  Pavements i2mo,  2  oo 

7 


Thurston's  Materials  of  Engineering.     3  Parts 8vo,  8  oo 

Part  I. — Non-metallic  Materials  of  Engineering  and  Metallurgy 8vo,  2  oo 

Part  II. — Iron  and  Steel 8vo,  3  50 

Part  III. — A  Treatise  on  Brasses,  Bronzes,  and  Other  Alloys  and  their 

Constituents 8vo,  a  50 

rhurston's  Text-book  of  the  Materials  of  Construction 8vo,  5  oo 

Tillson's  Street  Pavements  and  Paving  Materials 8vo,  4  oo 

Waddell's  De  Pontibus      (A  Pocket-book  for  Bridge  Engineers.). .  1 6mo,  mor.,  3  oo 

Specifications  for  Steel  Bridges I2mo,  i  23 

Wood's  Treatise  on  the  Resistance  of  Materials,  and  an  Appendix  on  the  Pres- 
ervation of  Timber   8vo,  a  oo 

Elements  of  Analytical  Mechanics 8vo,  3  oo 

Wood's  Rustless  Coatings.     (Shortly.) 


RAILWAY  ENGINEERING. 

Andre ws's  Handbook  for  Street  Railway  Engineers.     3X5  inches,  morocco,  i  25 

Berg's  Buildings  and  Structures  of  American  Railroads 4to,  5  oo 

Brooks's  Handbook  of  Street  Railroad  Location i6mo.  morocco,  i  50 

Butts's  Civil  Engineer's  Field-book i6mo,  morocco,  2  50 

Crandall's  Transition  Curve i6mo,  morocco,  i  50 

Railway  and  Other  Earthwork  Tables 8vo,  i  50 

Dawson's  "Engineering"  and  Electric  Traction  Pocket-book.    i6mo,  morocco,  5  oo 

Dredge's  History  of  the  Pennsylvania  Railroad:    (1879) Paper,  5  oo 

*  Drinker's  Tunneling,  Explosive  Compounds,  and  Rock  Drills,  4to,  half  mor.,    25  oo 

Fisher's  Table  of  Cubic  Yards Cardboard  25 

Godwin's  Railroad  Engineers'  Field-book  and  Explorers'  Guide i6mo,  mor.,  2   50 

Howard's  Transition  Curve  Field-book i6mo,  morocco,  i  50 

Hudson's  Tables  for  Calculating  the  Cubic  Contents  of  Excavations  and  Em- 
bankments    8vo,  i   oo 

Molitor  and  Beard's  Manual  for  Resident  Engineers i6mo,  i   oo 

Nagle's  Field  Manual  for  Railroad  Engineers i6mo,  morocco.  3  oo 

Philbrick's  Field  Manual  for  Engineers i6mo,  morocco,  3  oo 

Pratt  and  Alden's  Street-railway  Road-bed 8vo,  2  oo 

Searles's  Field  Engineering i6mo,  morocco,  3  oo 

Railroad  Spiral i6mo,  morocco,  i   50 

Taylor's  Prismoidal  Formulae  and  Earthwork 8vo,  i  50 

*  Trautwine's  Method  of  Calculating  the  Cubic  Contents  of  Excavations  and 

Embankments  by  the  Aid  of  Diagrams 8vo,  2  oo 

The  Field  Practice  of  paying    Out    Circular    Curves    for    Railroads. 

i2mo,  morocco,  2   50 

*  Cross-section  Sheet Paper,  25 

Webb's  Railroad  Construction.     2d  Edition,  Rewritten i6mn.  morocco,  5  oo 

Wellington's  Economic  Theory  of  the  Location  of  Railways Small  8vo,  5  oo 


DRAWING. 

Barr's  Kinematics  of  Machinery 8vo,  2  50 

*  Bartlett's  Mechanical  Drawing 8vo,  3  oo 

Abridged  Ed 8vo,  i  50 

Coolidge's  Manual  of  Drawing 8vo,  paper,  i  oo 

Durley's  Kinematics  of  Machines 8vo,  4  oo 

Hill's  Text-book  on  Shades  and  Shadows,  and  Perspective 8vo,  2  oo 

Jones's  Machine  Design: 

Part  I. — Kinematics  of  Machinery 8vo,  i  50 

Part  II. — Form,  Strength,  and  Proportions  of  Parts 8vo,  3  oo 

8 


MacCord's  Elements  of  Descriptive  Geometry 8vo,  3  oo 

Kinematics;  or,  Practical  Mechanism 8vo,  5  oo 

Mechanical  Drawing 4to,  4  oo 

Velocity  Diagrams 8vo,  I  50 

*  Mahan's  Descriptive  Geometry  and  Stone-cutting 8vo,  i  50 

Industrial  Drawing.    (Thompson.) 8vo,  3  50 

Reed's  Topographical  Drawing  and  Sketching 4to,  5  oo 

Reid's  Course  in  Mechanical  Drawing 8vo,  2  oo 

Text-book  of  Mechanical  Drawing  and  Elementary  Machine  Design.  .8 vo.  3  oo 

Robinson's  Principles  of  Mechanism 8vo,  3  oo 

Smith's  Manual  of  Topographical  Drawing.     (McMillan.) 8vo,  2  50 

Warren's  Elements  of  Plane  and  Solid  Free-hand  Geometrical  Drawing . .  I2mo,  oo 

Drafting  Instruments  and  Operations. . I2mo,  25 

Manual  of  Elementary  Projection  Drawing I2mo,  50 

Manual  of  Elementary  Problems  in  the  Linear  Perspective  of  Form  and 

Shadow I2mo,  oo 

Plane  Problems  in  Elementary  Geometry i2mo,  25 

Primary  Geometry I2mo,  75 

Elements  of  Descriptive  Geometry,  Shadows,  and  Perspective 8vo,  3  50 

General  Problems  of  Shades  and  Shadows : 8vo,  3  oo 

Elements  of  Machine  Construction  and  Drawing 8vo,  7  So 

Problems.  Theorems,  and  Examples  in  Descriptive  Geometrv 8vo,  2  50 

Weisbach's  Kinematics  and  the  Power  of  Transmission.       (Hermann  and 

Klein.) 8vo,  5  oo 

Whelpley's  Practical  Instruction  in  the  Art  of  Letter  Engraving i2mo,  a  oo 

Wilson's  Topographic  Surveying 8vo,  3  50 

Free-hand  Perspective 8vo,  2  50 

Free-hand  Lettering 8vo,  I  o« 

Woolf's  Elementary  Course  in  Descriptive  Geometry Large  8vo,  3  oo 

ELECTRICITY  AND   PHYSICS. 

Anthony  and  Brackett's  Text-book  of  Physics.     (Magie.) Small  8vo,  3  oo 

Anthony's  Lecture-notes  on  the  Theory  of  Electrical  Measurements 1 2mo,  i  oo 

Benjamin's  History  of  Electricity 8vo,  3  oo 

Voltaic  Cell 8vo,  3  oo 

Classen's  Quantitative  Chemical  Analysis  by  Electrolysis.    (Boltwood.).  .8vo,  3  oo 

Crehore  and  Souier's  Polarizing  Photo-chronograph 8vo,  3  oo 

Dawson's  "Engineering"  and  Electric  Traction  Pocket-book. .  i6mo,  morocco,  5  oo 
Dolezalek's    Theory    of    the    Lead    Accumulator.     (Storage  Battery.) 
(Shortly.)     (Von  Ende.) 

Duhem's  Thermodynamics  and  Chemistry.     (Burgess.) 8vo,  4  oo 

Flather's  Dvnamo  meters,  and  the  Measurement  of  Power 12010,  3  oo 

Giioert's  De  Magnete.     (Mottelay.) 8vo,  2  50 

Hanchett's  Alternating  Currents  Explained.     (Shortly.) 

Holman's  Precision  of  Measurements 8vo,  a  oo 

Telescopic  Mirror-scale  Method,  Adjustments,  and  Tests. Large  8vo,  75 

Lanaauer's  Spectrum  Analysis.    (Tingle.) 8vo,  3  oo 

Le  Chatelier's  High-temperature  Measurements.  (Boudouard — Burgess.  )i2mo,  3  oo 

LSb's  Electrolysis  and  Electrosynthesis  of  Organic  Compounds.  (Lorenz.)  i2mo,  i  oo 

*  Lyons's  Treatise  on  Electromagnetic  Phenomena.     Vols.  I.  and  n.  8vo,  each,  6  oo 

*  Michie.     Elements  of  Wave  Motion  Relating  to  Sound  and  Light 8vo,  4  oo 

Niaudet's  Elementary  Treatise  on  Electric  Batteries.     (Fishoack. ) i2mo,  2  50 

*  Parshall  and  Hobart's  Electric  Generators Small  4to.  half  morocco,  10  oo 

*  Rosenberg's  Electrical  Engineering.    (Haldane  Gee — Kinzbrunner.) 8vo,  i   50 

Ryan,  Jforris,  and  Hoxie's  Electrical  Machinery.     VoL  1 8vo,  2  56 

Thurston's  Stationary  Steam-engines 8vo,  2"  50 

*  Tillman's  Elementary  Lessons  in  Heat 8vo,  i  50 

9 


Tory  and  Pitcher's  Manual  of  Laboratory  Physics Small  8vo,  2  oo 

Hike's  Modern  Electrolytic  Copper  Refining 8vo,  3  oo 

LAW. 

*  Davis's  Elements  of  Law 8vo,  2  50 

*  Treatise  on  the  Military  Law  of  United  States 8vo,  7  oo 

*  Sheep,  7  50 

Manual  for  Courts-martial i6mo,  morocco,  i  50 

Wait's  Engineering  and  Architectural  Jurisprudence 8vo,  6  oo 

Sheep,  6  50 

Law  of  Operations  Preliminary  to  Construction  in  Engineering  and  Archi- 
tecture      8vo,  5  oo 

Sheep,  5  50 

Law  of  Contracts i 8vo,  3  oo 

Winthrop's  Abridgment  of  Military  Law i2mo,  2  50 

MANUFACTURES. 

Bernadou's  Smokeless  Powder — Nitro-cellulose  and  Theory  of  the  Cellulose 

Molecule i2mo,  2  50 

Holland's  Iron  Founder I2mo,  2  50 

"  The  Iron  Founder,"  Supplement i2mo,  2  50 

Encyclopedia  of  Founding  and  Dictionary  of  Foundry  Terms  Used  in  the 

Practice  of  Moulding I2mo,  3  oo 

Eissler's  Modern  High  Explosives 8vo,  4  oo 

Eff rent's  Enzymes  and  their  Applications.     (Prescott.) 8vo,  3  oo 

Fitzgerald's  Boston  Machinist i8mo,  i  oo 

Ford's  Boiler  Making  for  Boiler  Makers i8mo,  i  oo 

Hopkins's  Oil-chemists'  Handbook 8vo,  3  oo 

Keep's  Cast  Iron. 8vo,  2  50 

Leach's  The  Inspection  and  Analysis  of  Food  with  Special  Reference  to  State 

ControL     (In  preparation.) 

Metcalf's  SteeL     A  Manual  for  Steel-users i2mo,  a  oo 

Metcalfe's  Cost  of  Manufactures —And  the  Administration    of  Workshops, 

Public  and  Private 8vo,  5  oo 

Meyer's  Modern  Locomotive  Construction 4to,  10  oo 

*  Reisig's  Guide  to  Piece-dyeing 8vo,  25  oo 

Smith's  Press-working  of  Metals 8vo,  3  oo 

Wire :   Its  Use  and  Manufacture Small  4to,  3  oo 

Spalding's  Hydraulic  Cement I2mo,  2  oo 

Spencer's  Handbook  for  Chemists  of  Beet-sugar  Houses i6mo,  morocco,  3  oo 

^andboox  tor  sugar  Manutacturers  ana  their  Chemists..  .  i6mo,  morocco,  2  oo 
Thurston's  Manual  of  Steam-boilers,  their  Designs,  Construction  and  Opera- 
tion   8vo,  5  oo 

*  Walke's  Lectures  on  Explosives 8vo,  4  oo 

West's  American  Foundry  Practice i2mo,  2  50 

Moulder's  Text-book I2mo.  2  50 

Wiechmann's  Sugar  Analysis Small  8vo,  2  50 

Wolff's  Windmill  as  a  Prime  Mover 8vo,  3  oo 

Woodbury's  Fire  Protection  of  Mills 8vo,  2  50 

MATHEMATICS. 

Baker's  Elliptic  Functions 8vo,  i  50 

*  Bass's  Elements  of  Differential  Calculus * I2mo,  4  oo 

Briggs's  Elements  of  Plane  Analytic  Geometry i2mo,  i  oo 

10 


50 
50 
50 

25 

75 
50 


Compton's  Manual  of  Logarithmic  Computations i2mo, 

Davis's  Introduction  to  the  Logic  of  Algebra 8vo, 

*  Dickson's  College  Algebra   Large  i2mo, 

*  Introduction  to  the  Theory  of  Algebraic  Equations   Large  i2mo, 

Halsted's  Elements  of  Geometry    8vo, 

Elementary  Synthetic  Geometry 8vo. 

Rational  Geometry.     ( Shortly. ) 

*  Johnson's  Three-place  Logarithmic  Tables:    Vest-pocket  size paper,        15 

100  copies  for  5  oo 

*  Mounted  on  heavy  cardboard,  8  X 10  inches,         25 

10  copies  for  2  oo 

Elementary  Treatise  on  the  Integral  Calculus Small  8vo,  i  50 

Curve  Tracing  in  Cartesian  Co-ordinates I2mo,  i  oo 

Treatise  on  Ordinary  and  Partial  Differential  Equations Small  8vo,  3  SO 

Theory  of  Errors  and  the  Method  of  Least  Squares I2mo,  i  50 

*  Theoretical  Mechanics i2mo,  3  oo 

Laplace's  Philosophical  Essay  on  Probabilities.     (Truscott  and  Emory.)  i2mo,  2  oo 

*  Ludlow  and  Bass.     Elements  of  Trigonometry  and  Logarithmic  and  Other 

Tables 8vo,  3  oo 

Trigonometry  and  Tables  published  separately Each,  2  oo 

Maurer's  Technical  Mechanics 8vo,  4  oo 

Merriman  and  Woodward's  Higher  Mathematics 8vo,  5  oo 

Merriman's  Method  of  Least  Squares 8vo,  2  oo 

Rice  and  Johnson's  Elementary  Treatise  on  the  Differential  Calculus .  Sm.,  8vo,  3  oo 

Differential  and  Integral  Calculus.     2  vols.  in  one Small  8vo,  2  50 

Wood's  Elements  of  Co-ordinate  Geometry 8vo,  2  oo 

Trigonometry:  Analytical,  Plane,  and  Spherical i2mo,  i  oo 

MECHANICAL   ENGINEERING. 
MATERIALS  OF  ENGINEERING,  STEAM-ENGINES  AND  BOILERS. 

Baldwin's  Steam  Heating  for  Buildings I2mo,  2  50 

Barr's  Kinematics  of  Machinery 8vo,  2  50 

*  Bartlett's  Mechanical  Drawing 8vo,  3  oo 

*  "  "  "        Abridged  Ed 8vo,  i  50 

Benjamin's  Wrinkles  and  Recipes i2mo,  2  oo 

Carpenter's  Experimental  Engineering .8vo,  6  oo 

Heating  and  Ventilating  Buildings 8vo,  4  oo 

Gary's  Smoke  Suppression  in  Plants  using  Bituminous  Coal.      (In  prep- 
aration. ) 

Clerk's  Gas  and  Oil  Engine Small  8vo,  4  oo 

Coolidge's  Manual  of  Drawing 8vo,    paper,  i  oo 

Cromwell's  Treatise  on  Toothed  Gearing i2mo,  i  50 

Treatise  on  Belts  and  PuLeys i2mo,  i  50 

Durley's  Kinematics  of  Machines 8vo,  4  oo 

Flather's  Dynamometers  and  the  Measurement  of  Power i2mo,  3  oo 

Rope  Driving i2mo,  2  oo 

Gill's  Gas  and  Fuel  Analysis  for  Engineers iamo,  I   25 

Hall's  Car  Lubrication. i  2mo,  i  oo 

Button's  The  Gas  Engine 8vo,  5  oo 

Jones's  Machine  Design: 

Part   I. — Kinematics  of  Machinery    8vo,  i  50 

Part  II. — Form,  Strength,  and  Proportions  of  Parts 8vo,    3  oo 

Kent's  Mechanical  Engineer's  Pocket-book i6mo,    morocco,  5  oo 

Kerr's  Power  and  Power  Transmission 8vo,  2  oo 

MacCord's  Kinematics;  or,  Practical  Mechanism 8vo,    5  oo 

Mechanical  Drawing 4to,    4  oo 

Velocity  Diagrams 8vo,    150 

11 


Mahan's  Industrial  Drawing.    (Thompson.) 8vo,  3  50 

Poole's  Calorific  Power  of  Fuels 8vo,  3  oo 

Reid's  Course  in  Mechanical  Drawing 8vo.  2  oo 

Text-book  of  Mechanical  Drawing  and  Elementary  Machine  Design.  .8vo,  3  oo 

Richards's  Compressed  Air i2mo,  i  50 

Robinson's  Principles  of  Mechanism 8vo,  3  oo 

Smith's  Press-working  of  Metals 8vo,  3  oo 

Thurston's  Treatise  on   Friction  and    Lost  Work  in    Machinery  and   Mul 

Work 8vo,  3  oo 

Animal  as  a  Machine  and  Prime  Motor,  and  the  Laws  of  Energetics .  121110,  i  oo 

Warren's  Elements  of  Machine  Construction  and  Drawing Svo,  750 

Weisbach's  Kinematics  and  the  Power  of  Trarsmission.      Herrmann- 
Klein.) 8vo,  5  oo 

Machinery  of  Transmission  and  Governors.     (Herrmann — Klein.).  .8vo,  5  oo 

Hydraulics  and  Hydraulic  Motors.     (Du  Bois.) 8vo,  5  oo 

Wolff's  Windmill  as  a  Prime  Mover 8vo,  3  oo 

Wood's  Turbines ...                                                                                      ...  8vo,  2  50 


MATERIALS  OF  ENGINEERING. 

Bovey's  Strength  of  Materials  and  Theory  of  Structures 8vo,  7  50 

Burr's  Elasticity  and  Resistance  of  the  Materials  of  Engineering.     6th  Edition, 

Reset 8vo.  7  so 

Church's  Mechanics  of  Engineering 8vo,  6  oo 

Johnson'«s  Materials  of  Construction Large  8vo,  6  oo 

Keep's  Cast  Iron 8vo,  2  50 

Lanza's  Applied  Mechanics 8vo,  7  So 

Martens's  Handbook  on  Testing  Materials.     (Henning.) 8vo,  7  So 

Merriman's  Text-book  on  the  Mechanics  of  Materials 8vo,  4  oo 

Strength  of  Materials i2mo,  i  oo 

Metcalf's  SteeL     A  Manual  for  Steel-users i2mo  2  oo 

Smith's  Wire:  Its  Use  and  Manufacture Small  410,  3  oo 

Materials  of  Machines i2mo  i  oo 

Thurston's  Materials  of  Engineering 3  vols  ,  Svo,  8  oo 

Part   H.— Iron  and  Steel Svo,  3  50 

Part  IH.— A  Treatise  on  Brasses,  Bronzes,  and  Other  Alloys  and  their 

Constituents Svo  2  50 

Text-book  of  the  Materials  of  Construction Svo,  5  oo 

Wood's  Treatise  on  the  Resistance  of  Materials  and  an  Appendix  on  the 

Preservation  of  Timber Svo,  2  oo 

Elements  of  Analytical  Mechanics Svo,-  3  oo 

Wood's  Rustless  Coatings.     ( Shortly. ) 


STEAM-ENGINES  AND  BOILERS. 

Carnot's  Reflections  on  the  Motive  Power  of  Heat.     (Thurston.) i2mo,    l   50 

Dawson's  "Engineering"  and  Electric  Traction  Pocket-book. .  i6mo,  mor.,    5  oo 

Ford's  Boiler  Making  for  Boiler  Makers iSmo,    i  oo 

Goss's  Locomotive  Sparks Svo,    2  oo 

Hemimway's  Indicator  Practice  and  Steam-engme  Economy i2mo,    2  oo 

Button's  Mechanical  Engineering  of  Power  Plants Svo,    5  oo 

Heat  and  Heat-engines Svo, 

Kent's  Steam-bo'ler  Economy Svo, 

Kneass's  Practice  and  Theory  of  the  Injector Svo 

MacCord's  Slide-valves Svo, 

Meyer's  Modern  Locomotive  Construction 4to. 

12 


Peabody's  Manua,  of  the  Steam-engine  Indicator i2mo,  i  50 

Tables  of  the  Properties  of  Saturated  Steam  and  Other  Vapors 8vo,  i  oo 

Thermodynamics  of  the  Steam-engine  and  Other  Heat-engines 8vo,  5  oo 

Valve-gears  for  Steam-engines 8vo,  2  50 

Peabody  and  Miller's  Steam-boilers 8vo,  4  oo 

Pray's  Twenty  Years  with  the  Indicator Large  8vo,  2  50 

Pupln's  Thermodynamics  of  Reversible  Cycles  in  Gases  and  Saturated  Vapors. 

(Osterberg.) i2mo.  i  25 

Reagan's  Locomotives :  Simple,  Compound,  and  Electric 1 2010,  2  50 

Rontgen's  Principles  of  Thermodynamics.     (Du  Bois.) 8vo,  5  oo 

Sinclair's  Locomotive  Engine  Running  and  Management i2mo,  2  oo 

Smart's  Handbook  of  Engineering  Laboratory  Practice i2mo,  2  50 

Snow's  Steam-boiler  Practice 8vo,  3  oo 

Spangler's  Valve-gears 8vo,  2  50 

Notes  on  Thermodynamics I2mo,  i  oo 

Spangler,  Greene,  and  Marshall's  Elements  of  Steam-engineering 8vo,  3  oo 

Thurston's  Handy  Tables 8vo.  i    50 

Manual  of  the  Steam-engine 2  vols.   8vo,  10  oo 

Part  I. — History.  Structuce,  and  Theory 8vo,  6  oo 

Part  H. — Design,  Construction,  and  Operation 8vo,  6  oo 

Handbook  of  Engine  and  Boiler  Trials,  and  the  Use  of  the  Indicator  and 

the  Prony  Brake 8vo^  5  oo 

Stationary  Steam-engines ; 8vo,  2  50 

Steam-boiler  Explosions  in  Theory  and  in  Practice I2mo  i   50 

Manual  of  Steam-boilers  ,  Their  Designs,  Construction,  and  Operation .  8vo,  5  oo 

Weisbach's  Heat,  Steam,  a    J  Steam-engines.     (Du  Bois.) 8vo,  5  oo 

Whitham's  Steam-engine  1  isign 8vo,  5  oo 

Wilson's  Treatise  on  Steam-boilers.     (Flather.) i6mo,  2  50 

Wood's  Thermodynamics.  Heat  Motors,  and  Refrigerating  Machines.  .  .  .8vo,  4  oo 


MECHANICS    AND  MACHINERY. 

Barr's  Kinematics  ot  Machinery. 8vo,  2  50 

Bovey's  Strength  of  Materials  and  Theory  of  Structures 8vo,  7  50 

Chase's  The  Art  of  Pattern-making i2mo,  2  50 

Chordal. — Extracts  from  Letters   I2mo,  2  oo 

Church's  Mechanics  of  Engineering 8vo,  6  oo 

Notes  and  Examples  in  Mechanics 8vo,  oo 

Compton's  First  Lessons  in  Metal-working I2mo,  50 

Compton  and  De  Groodt's  The  Speed  Lathe i2mo,  50 

Cromwell's  Treatise  on  Toothed  Gearing   i2mo,  50 

Treatise  on  Belts  and  Pulleys i2mo,  50 

Dana's  Text-book  of  Elementary  Mechanics  for  the   Use  of  Colleges  and 

Schools i2mo,  i  50 

Dingey's  Machinery  Pattern  Making. 12010,  2  oo 

Dredge's   Record  of  the   Transportation   Exhibits  Building  of  the   World's 

Columbian  Exposition  of  1893   4to,  half  morocco,  5  oo 

Du  Bois's  Elementary  Principles  of  Mechanics : 

VoL     I. — Kinematics 8vo,  3  50 

Vol    II.— Statics 8vo,  4  oo 

Vol.  III.— Kinetics 8vo,  3  so 

Mechanics  of  Engineering.     VoL    I Small  4to,  7  50 

Vol.  II Small  4to,  10  oo 

Durley's  Kinematics  of  Machines 8vo,  4  oo 

Fitzgerald's  Boston  Machinist i6mo.  i   oo 

Flather's  Dynamometers,  and  the  Measurement  of  Power i2mo,  3  oo 

Rope  Driving I2mo,  2  oo 

Goss's  Locomotive  Sparks 8vo,  2  oo 

13 


Hall's  Car  Lubrication 

Holly's  Art  of  Saw^Filing i8mo 

*  Johnson's  Theoretical  Mechanics I2mo,  3 

Statics  by  Graphic  and  Algebraic  Methods 8vo,  2 

Jones's  Machine  Design: 

Part   I. — Kinematics  of  Machinery 8vo,  i 

Part  II. — Form,  Strength,  and  Proportions  of  Parts 8vo,  3 

Kerr's  Power  and  Power  Transmission 8vo,  2 

Lanza's  Applied  Mechanics 8vo,  7 

MacCord's  Kinematics;  or,  Practical  Mechanism 8^0,  5 

Velocity  Diagrams 8vo ,  i 

Maurer's  Technical  Mechanics 8vo,  4 

Merriman's  Text-book  on  the  Mechanics  of  Materials 8vo,  4 

*  Michie's  Elements  of  Analytical  Mechanics   8vo.  4 

Reagan's  Locomotives:  Simple,  Compound,  and  Electric i2mo,  2 

Reid's  Course  in  Mechanical  Drawing 8vo , 

Text-book  of  Mechanical  Drawing  and  Elementary  Machine  Design.  .8vo, 

Richards's  Compressed  Air i2mo,  i   50 

Robinson's  Principles  of  Mechanism 8vo,  3  oo 

Ryan,  Norris,  and  Hoxie's  Electrical  Machinery 8vo,  2  50 

Sinclair's  Locomotive-engine  Running  and  Management I2mo,  2  oo 

Smith's  Press-working  of  Metals 8vo,  3  oo 

Materials  of  Machines i2mo,  i  oo 

Spangler,  Greene,  and  Marshall's  Elements  of  Steam-engineering 8vo,  3  oo 

Thurston's  Treatise  on  Friction  and  Lost  Work  in  Machinery  and   Mill 

Work 8vo,  3  oo 

Animal  as  a  Machine  and  Prime  Motor,  and  the  Laws  of  Energetics.  i2mo,  i  oo 

Warren's  Elements  of  Machine  Construction  and  Drawing 8vo,  7  So 

Weisbach's    Kinematics    and    the   Power  of    Transmission.     (Herrmann — 
Klein.) 8vo, 

Machinery  of  Transmission  and  Governors.     (Herrmann — Klein.). 8vo, 
Wood's  Elements  of  Analytical  Mechanics 8vo, 

Principles  of  Elementary  Mechanics i2mo, 

Turbines 8vo,  2 

The  World's  Columbian  Exposition  of  1893 4to,  i 

METALLURGY. 

Egleston's  Metallurgy  of  Silver,  Gold,  and  Mercury: 

VoL   I.— Silver 8vo, 

Vol.   II.— Gold  and  Mercury 8vo, 

**  Iles's  Lead-smelting.     (Postage  9  cents  additional.) i2mo, 

Keep's  Cast  Iron 8vo, 

Kunhardt's  Practice  of  Ore  Dressing  in  Europe 8vo, 

Le  Chatelier's  High-temperature  Measurements.   (Boudouard — Burgess.) .  i2mo,  3  oo 

Metcalf 's  Steel.     A  Manual  for  Steel-users I2mo,    2  oo 

Smith's  Materials  of  Machines i2mo,    i   oo 

Thurston's  Materials  of  Engineering.     In  Three  Parts 8vo,    8  oo 

Part   II.— Iron  and  Steel 8vo,    3  5<> 

Part  III. — A  Treatise  on  Brasses,  Bronzes,  and  Other  Alloys  and   their 

Constituents 8vo,    2  50 

Ulke's  Modern  Electrolytic  Copper  Refining. 8vo,    3  oo 

MINERALOGY. 

Barringer's  Description  of  Minerals  of  Commercial  Value.     Oblong,  morocco,  2  50 

Boyd's  Resources  of  Southwest  Virginia 8vo,  3  oo 

Map  of  Southwest  Virginia Pocket-book  form,  2  oo 

14 


finish's  Manual  of  Determinative  Mineralogy.     (Penfield.) 8vo,  4  oo 

Chester's  Catalogue  of  Minerals 8vo,  paper,  i  oo 

Cloth,  i  25 

Dictionary  of  the  Names  of  Minerals 8vo,  3  50 

Dana's  System  of  Mineralogy Large  8vo,  half  leather,  12  50 

First  Appendix  to  Dana's  New  "System  of  Mineralogy.". . .  .Large  8vo,  i  oo 

Text-book  of  Mineralogy 8vo,  4  oo 

Minerals  and  How  to  Study  Them. . . : i2mo,  i  50 

Catalogue  of  American  Localities  of  Minerals Large  8vo,  i  oo 

Manual  of  Mineralogy  and  Petrography i2mo,  2  oo 

Eakle's  Mineral  Tables.     (Shortly.) 

Egleston's  Catalogue  of  Minerals  and  Synonyms 8vo,  2  50 

Hussak's  The  Determination  of  Rock-forming  Minerals.     (Smith.)  Small  8vo,  2  oo 

Merrill's  Non- Metallic  Minerals.     (Shortly.) 

*  Penfield's  Notes  on  Determinative  Mineralogy  and  Record  of  Mineral  Tests. 

8vo,  paper,  o  50 
Rosenbusch's   Microscopical   Physiography   of   the    Rock-making   Minerals. 

(Iddings.) 8vo,  5  oo 

*  Tollman's  Text-book  of  Important  Minerals  and  Docks 8vo,  2  oo 

Williams's  Manual  of  Lithology 8vo,  3  oo 

MINING. 

Beard's  Ventilation  of  Mines i2mo,  2  50 

Boyd's  Resources  of  Southwest  Virginia 8vo,  3  oo 

Map  of  Southwest  Virginia Pocket-book  form,  2  oo 

*  Drinker's  Tunneling,  Explosive  Compounds,  and  Rock  Drills. 

4to,  half  morocco,  25  oo 

Eissler's  Modern  High  Explosives 8vo,  4  oo 

Fowler's  Sewage  Works  Analyses I2mo,  2  oo 

Goodyear's  Coal-mines  of  the  Western  Coast  of  the  United  States i2mo,  2  50 

Ihlseng's  Manual  of  Mining 8vo,  4  oo 


**  Iles's  Lead-smelting.     (Postage  QC.  additional.) I2mo, 

Kunhardt's  Practice  of  Ore  Dressing  in  Europe 8vo, 

O'Driscoll's  Notes  on  the  Treatment  of  Gold  Ores 8vo, 

*  Walke's  Lectures  on  Explosives 8vo, 

Wilson's  Cyanide  Processes I2mo, 

Chlorination  Process i2mo, 

Hydraulic  and  Placer  Mining i2mo, 

Treatise  on  Practical  and  Theoretical  Mine  Ventilation i2mo 


50 
50 
oo 

00 

50 
50 

00 
25 


SANITARY  SCIENCE. 

Copeland's  Manual  of  Bacteriology.     (In  preparation.) 

FolwelTs  Sewerage.     (Designing,  Construction  and  Maintenance.; 8vo,  3  oo 

Water-supply  Engineering 8vo,  4  oo 

Fuertes's  Water  and  Public  Health I2mo,  i  50 

Water-filtration    Works I2mo,  2  50 

Gerhard's  Guide  to  Sanitary  House-inspection i6mo,  i  oo 

Goodrich's  Economical  Disposal  of  Town's  Refuse Demy  8vo,  3  50 

Hazen's  Filtration  of  Public  Water-supplies 8vo,  3  oo 

Kiersted's  Sewage  Disposal I2mo,  i  25 

Leach's  The  Inspection  and  Analysis  of  Food  with  Special  Reference  to  State 

Control.     (In  preparation.) 

Mason's    Water-supply.     (Considered    Principally   from   a    Sanitary    Stand- 
point.)    3d  Edition,  Rewritten 8vo,  4  oo 

Examination  of  Water.     (Chemical  and  Bacteriological.) 12 mo,  i  25 

15 


Merriman's  Elements  of  Sanitary  Engineering 8vo,    2  oo 

Nichols's  Water-supply.     (Considered  Mainly  from  a  Chemical  and  Sanitary 

Standpoint.)     (1883.) 8vo, 

Ogden's  Sewer  Design I2mo, 

*  Price's  Handbook  on  Sanitation. .'. I2mo, 

Richards's  Cost  of  Food.     A  Study  in  Dietaries , i2mo, 


Cost  of  Living  as  Modified  by  Sanitary  Science i2mo, 


5o 
oo 
So 

00 
00 


Richards  and  Woodman's  Air,  Water,  and  Food  from  a  Sanitary  Stand- 
point   8vo,  2  oo 

*  Richards  and  Williams's  The  Dietary  Computer 8vo,    i  50 

Rideal's  Sewage  and  Bacterial  Purification  of  Sewage 8vo,  3  50 

Turneaure  and  Russell's  Public  Water-supplies 8vo,  5  oo 

Whipple's  Microscopy  of  Drinking-water 8vo,  3  SO 

WoodhulTs  Notes  and  Military  Hygiene i6mo,  i  50 

MISCELLANEOUS. 

Barker's  Deep-sea  Soundings 8vo,  2  oo 

Emmons's  Geological  Guide-book  of  the  Rdtky  Mountain  Excursion  of  the 

International  Congress  of  Geologists Large  8vo,  i  50 

FerrePs  Popular  Treatise  on  the  Winds. 8vo,  4  oo 

Haines's  American  Railway  Management I2mo,  2  go 

Mott's  Composition,  Digestibility,  and  Nutritive  Value  of  Food.   Mounted  chart,  i  25 

Fallacy  of  the  Present  Theory  of  Sound i6mo,  i  oo 

Ricketts's  History  of  Rensselaer  Polytechnic  Institute,  1824-1894.  Small  8vo,  3  oo 

Rotherham's  Empnasized  New  Testament Large  8vo,  2  oo 

Steel's  Treatise  on  the  Diseases  of  the  Dog 8vo,  3  50 

Totten's  Important  Question  in  Metrology 8vo,  2  50 

The  World's  Columbian  Exposition  ot  1893 4to,  i  oo 

Worcester  and  Atkinson.     Small  Hospitals,  Establishment  and  Maintenance, 
and  Suggestions  for  Hospital  Architecture,  with  Plans  for  a  Small 

Hospital i2mo,  i   25 

HEBREW  AND  CHALDEE  TEXT-BOOKS. 

Green's  Grammar  of  the  Hebrew  Language 8vo,  3  00 

Elementary  Hebrew  Grammar i2mo,  i  25 

Hebrew  Chrestomathy   8vo,  2  oo 

Gesenius's  Hebrew  and  Chaldee  Lexicon  to  the  Old  Testament  Scriptures. 

(Tregelles.) Small  4to,  half  morocco,  5 j »o 

Letteris's  Hebrew  Bible 8vo,  2  25 

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